Substituted Hydrophobic Benzene Sulfonamide Thiazole Compounds for Use in Treating Cancer

ABSTRACT

The present invention relates to compound of general formula (I) R 1  is selected from H, aryl and alkyl, R 2  is selected from H, alkyl, aryl and CO—R 6 ; R 3  is selected from H, halogen, alkyl, alkenyl, alkynyl, aryl, NHR 7 , NR 7 R 8 , OR 7  and SR 7 ; R 4  is selected from (C 6 -C 12 ) alkyl, (C 2 -C 12 ) alkenyl, (C 2 -C 12 ) alkynyl and (C 6 -C 10 ) aryl, R 5  represents H, R 6 , aryl, OH, OR 6 , O-aryl, SH, SR 6 , S-aryl, CN, NO 2 , CF 3 , COOR 6 , SO 2 NR 6 R 7 , CONR 6 R 7 , NH 2 , NHR 6 , NH-aryl, NR 6 R 7 , NHCOR 6  or aminoacyl; R 6  is alkyl optionally substituted with halogen, OH, SH, NH 2 , O-alkyl, S-alkyl, NH-alkyl or NH-di(alkyl); R 7  and R 8  identical or different are H or alkyl optionally substituted with halogen, OH, SH, NH 2 , O-alkyl, S-alkyl, NH-alkyl or NH-di (alkyl), their pharmaceutically acceptable salts and/or isomers, tautomers, solvates or isotopic variations thereof. The compounds are useful for the treatment of cancers.

FIELD OF THE INVENTION

The invention relates to new benzene sulfonamide thiazole compoundsactive for the treatment of cancers.

BACKGROUND OF THE INVENTION

Cutaneous melanoma deriving from the transformation of melanocytes isone of the most lethal cancers among young adults. Its incidence hasincreased at a dramatic rate during the last decades. Melanoma has ahigh capability of invasion and rapid metastasis to other organs. Theprognosis of metastatic melanoma is extremely pejorative, as the variousprotocols of chemotherapy or immunotherapy have not shown real survivalbenefit. Indeed, at the ganglionic metastatic stage, the forecastdeteriorates considerably with a survival rate after 5 years of 50%. Atthe stage of visceral metastases, the forecast is catastrophic with amedian of survival of 6 months. Therefore, the melanoma, whichrepresents only 5% of the cutaneous cancers, represents 80% of thedeaths associated to this type of cancer. With an incidence, whichdoubles every ten years (10000 new cases in France in 2007), themelanoma constitutes a real problem of public health. Finally, even ifrecently encouraging results were obtained with vemurafenib anddabrafenib, two inhibitors of the B-Raf pathway, the responses remaintransitory. Indeed, vemurafenib and dabrafenib target only melanomasmutated on B-Raf (approximately 50% of the metastatic melanomas).Unfortunately, after a short period of regression, the melanoma acquiresin all cases, a resistance against the drug and the metastases developagain, increasing only about 2 months the life expectancy of thepatient. The identification of these mechanisms of resistance is now thesubject matter of numerous works but no study managed to clearlyidentify the mechanisms involved.

Recently, the anti-CTLA4 antibody ipilimumab able to reactivate theimmune response of the patient was developed for the treatment ofmelanoma. However, this approach provides an objective response in only10 to 15% of the patients.

The identification of new candidate molecules is thus a major aim forthe development of specific biotherapies.

WO2014072486 describes a series of benzene sulfonamide thiazolecompounds invented by the instant inventors, which are active in thetreatment of cancer, especially on melanoma cells.

The inventors have now optimized the series described in WO2014072486and generated novel hydrophobic derivatives showing a substantiallyhigher potency in models of melanoma while having signaling pathways andmechanisms of action totally different from those of dabrafenib despitea structural similarity with dabrafenib.

In addition, it appears that the compounds of the invention are alsoefficient on several other cancers namely prostate, breast and colonindicating that these molecules may be active in all type of cancers.

SUMMARY OF THE INVENTION

The invention relates to benzene sulfonamide thiazole compounds ofgeneral formula:

-   -   in which R₁ to R₅ have the meanings indicated below, and to        processes for the preparation of, compositions containing and        the uses of such derivatives.

DETAILED DESCRIPTION OF THE INVENTION

The invention relates to compounds of general formula (I):

wherein

-   -   R₁ is selected from H, aryl and alkyl;    -   R₂ is selected from H, alkyl, aryl and CO—R₆;    -   R₃ is selected from H, halogen, alkyl, alkenyl, alkynyl, aryl,        NHR₇, NR₇R₈, OR₇ and SR₇;    -   R₄ is selected from (C₆-C₁₂) alkyl, (C₂-C₁₂) alkenyl, (C₂-C₁₂)        alkynyl and (C₆-C₁₀) aryl, wherein aryl is unsubstituted or        substituted by 1 to 5 alkyl groups, wherein the alkyl, alkenyl        and alkynyl groups are linear, branched or cyclic and wherein        the alkynyl group is optionally substituted with one to three OH        groups;    -   R₅ is selected from H, R₆, aryl, OH, OR₆, O-aryl, SH, SR₆,        S-aryl, CN, NO₂, CF₃, COOR₆, SO₂NR₇R₈, CONR₇R₈, NH₂, NHR₆,        NH-aryl, NR₇R₈, NHCOR₆ and aminoacyl;    -   R₆ is alkyl optionally substituted with halogen, OH, SH, NH₂,        O-alkyl, S-alkyl, NH-alkyl or NH-di(alkyl);    -   R₇ and R₈ identical or different are H or alkyl optionally        substituted with halogen, OH, SH, NH₂, O-alkyl, S-alkyl,        NH-alkyl or NH-di (alkyl).

The invention further pertains to compounds of formula (I) above and, ifappropriate, their pharmaceutically acceptable salts and/or isomers,tautomers, solvates or isotopic variations thereof for use in thetreatment of cancer.

In the above general formula (I), unless specified otherwise:

-   -   Alkyl denotes a straight-chain or branched group containing 1,        2, 3, 4, 5 or 6 carbon atoms. Examples of suitable alkyl        radicals are methyl, ethyl, n-propyl, isopropyl, n-butyl,        isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl,        etc.,    -   Cycloalkyl comprises 3 to 8 carbon atoms and includes        cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl        and cyclooctyl,    -   Alkenyl denotes a straight-chain or branched group containing 2,        3, 4, 5 or 6 carbon atoms and one or more double bonds. Examples        of suitable alkenyl radicals are, ethenyl, n-propenyl,        isopropenyl, n-butenyl, isobutenyl, sec-butenyl, tert-butenyl,        n-pentenyl, isopentenyl, n-hexenyl, etc.,    -   Alkynyl denotes a straight-chain or branched group containing 2,        3, 4, 5 or 6 carbon atoms and one or more triple bonds and        optionally one or more double bonds. Examples of suitable        alkenyl radicals are ethynyl, n-propynyl, isopropynyl,        n-butynyl, isobutynyl, sec-butynyl, tert-butynyl, n-pentynyl,        isopentynyl, n-hexynyl, etc.,    -   Aryl denotes an aromatic carbon ring comprising from 6 to 10        carbon atoms. A preferred aryl group is phenyl,    -   Aminoacyl means a group of formula RC(NH₂)COOH. Preferred groups        are those derived from glycine, lysine, tyrosine, arginine and        other natural or non-natural derivatives,    -   The free bond on the phenyl group means that the phenyl can be        substituted in the ortho, meta or para position.

In the above general formula (I), the following meanings are preferred:

-   -   R₁ is H,    -   R₂ is selected from H, methyl or COCH₃,    -   R₃ is H,    -   R₄ is (C₆-C₁₂) alkyl, preferably hexyl, heptyl, octyl, or    -   R₄ is (C₆-C₁₀ aryl), preferably phenyl or substituted phenyl, or    -   R₄ is CH═CHR₉, wherein R₉ is (C₁-C₈) alkyl, preferably hexyl, or    -   R₄ is C≡CR₁₀, wherein R₁₀ is selected from H, C₁-C₈ alkyl,        hydroxy (C₁-C₈) alkyl, cyclo (C₃-C₈) alkyl and hydroxyl-cyclo        (C₃-C₈ alkyl),    -   R₄ is in the meta or para position with respect to the sulfonyl        group,    -   R₅ is H,    -   R₆ is alkyl,    -   R₇ and R₈ are H.

Preferred compounds according to the invention are the following:

-   N-(4-(3-(4-(oct-1-ynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-(3-(oct-1-ynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-(3-(3-hydroxyprop-1-ynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-(3-((trimethylsilyl)ethynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-(3-ethynylphenylsulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(3-(2-aminothiazol-4-yl)phenyl)-4-(oct-1-ynyl)benzenesulfonamide-   N-(3-(2-(methylamino)thiazol-4-yl)phenyl)-4-(oct-1-ynyl)benzenesulfonamide-   4-(oct-1-ynyl)-N-(3-(2-(phenylamino)thiazol-4-yl)phenyl)benzenesulfonamide-   N-(4-(3-((4-(cyclohexylethynyl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-([1,1′-biphenyl]-4-sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-((2,3-dihydro-1H-indene)-5-sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-((4′-methyl-[1,1′-biphenyl])-4-sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-((4-octylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-((4-hexylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-((4-(3-hydroxyprop-1-yn-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide-   (Z)—N-(4-(3-((4-(oct-1-en-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(3-((4-ethynylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide.-   N-(4-(4-(4-methylpiperazin-1-yl)-3-((4-pentylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide-   N-(4-(4-morpholino-3-((4-pentylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide.

The compounds of the formula (I) may be prepared using conventionalprocedures such as by the following illustrative methods (schemes 1-2)in which the various substituents are as previously defined for thecompounds of the formula (I) unless otherwise stated.

The procedure for preparing the compounds of the invention according toscheme 1 comprises the following steps:

Step 1 to prepare compound of formula 2a: by bromination of compounds offormula 1a with a suitable brominating agent Br₂ or N-bromosuccinimide(NBS) in solvents such as ether (Et₂O), THF or MeTHF, preferably in thepresence of Lewis acid such as AlCl₃ (D. Guianvarch, R. Benhida, J-L.Fourrey, R. Maurisse, J-S. Sun. J. Chem. Soc. Chem. Comm. 2001,1814-1815).

Step 2 to prepare compound of formula 4a: this step consists ofcondensing compound 2a with thiourea of formula 3 in suitable solventsthat include but not restricted to EtOH, iPrOH, ethyl acetate, CH₂Cl₂,DMF. The reaction may be carried out at a temperature of about 25° C. to100° C., preferably at 60-80° C. with or without acid or base catalystdepending on the reactivity of the starting material.

Step 3 to prepare compounds of formula 5a: the reduction may be carriedout with a source of H₂ in the presence of metal catalyst which includebut not limited to palladium derivatives on carbon, platinum derivativeson carbon or Raney nickel on carbon or other source of H₂ such asNaBH₄/Pd/C, metal under acidic conditions (iron, tin chloride, titaniumchloride, Zinc in HCl or AcOH). The reaction could be realized in inertsolvents that include but are not restricted to EtOH, MeOH, THF,dioxane, AcOH, ethylacetate, at either atmospheric or elevated pressure.

Step 4 to prepare compounds of formula Ia: the reaction is typicallycarried out by reacting compounds of formula 5a with sulfonyl chlorideof formula 6a in an appropriate solvent such as CH₂Cl₂, AcOEt, DMF,DMSO, ether, THF, MeTHF, dioxane, acetonitrile in the presence of aminesuch as triethylamine, diisopropylethylamine, pyridine and substitutedpyridines (for example DMAP). The reaction may be also carried out inpyridine as solvent.

Step 5 to prepare compounds of formula Ha: the reaction is carried outby reacting compounds of formula Ia (X=halogen, preferably I, Br and Cl)with terminal alkynes, under Sonogashira conditions, typically in thepresence of palladium source, CuI, Et₃N, in an appropriate solvent suchas DMF, DMSO, THF, MeTHF, dioxane, acetonitrile

Step 6: reduction of the triple bond is performed by Lindlar-Pd—H2 orPd/C—H2

The procedure for preparing the compounds of the invention (compoundsIb, IIb and IIIb) according to scheme 2 comprises the following steps:

Step 1 to prepare compound of formula 2b: the carbon-carbon formationmay be achieved using techniques conventional in the art. In a typicalreaction, compound of formula 1b (X=leaving group in palladiumreactions, preferably Br or I) may be reacted with boron derivatives(Suzuki-Miyaura coupling, Palladium-Catalyzed Cross-Coupling Reactionsof Organoboron Compounds N. Miyaura, A. Suzuki Chem. Rev., 1995, 95 (7),pp 2457-2483), tin derivatives (Stille coupling, J. K. Stille, Angew.Chem. Int. Ed. Engl. 1986, 25, 508-524, D. Guianvarc'h, J-L Fourrey,J-S. Sun, R. Maurisse, R. Benhida. Bioorg. Med. Chem. 2003, 11,2751-2759) or by a direct C—H activation (J. Yamaguchi, A. D. Yamaguchi,K. Itami Angew. Chem. Int. Ed. 2012, 51, 8960-9009) in an appropriatesolvent for example as DMF, DMSO, THF, MeTHF, dioxane, acetonitrile, inthe presence of palladium catalyst for example Pd(PPh₃)₄, Pd(PPh₃)₂Cl₂,palladium dibenzylideneacetone at a temperature of 20 to 140° C.,preferably, 25-70° C. Depending on the nature of starting materials,this reaction requires some time other additives such as base(carbonate, amine) and/or ligands (phosphines) and/or copper source forexample CuI or other conventional additives in the art.

Steps 2, 3, 4 and 5 in scheme 2 are similar to those described above inscheme 1, e.g., step 3, 4, 5 and 6, respectively (reduction,sulfonylation and alkylation).

Pharmaceutically acceptable salts of the compounds of formula (I)include the acid addition and base salts thereof.

Suitable acid addition salts are formed from acids, which form non-toxicsalts. Examples include the acetate, aspartate, benzoate, besylate,bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, citrate,edisylate, esylate, formate, fumarate, gluceptate, gluconate,glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride,hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate,maleate, malonate, mesylate, methylsulphate, naphthylate, 2-napsylate,nicotinate, nitrate, orotate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate, stearate,succinate, tartrate, tosylate and trifluoroacetate and xinafoate salts.

Suitable base salts are formed from bases, which form non-toxic salts.Examples include the aluminium, arginine, benzathine, calcium, choline,diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine,potassium, sodium, tromethamine and zinc salts. Hemisalts of acids andbases may also be formed, for example, hemisulphate and hemicalciumsalts.

For a review on suitable salts, see “Handbook of Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH,Weinheim, Germany, 2002). Pharmaceutically acceptable salts of compoundsof formula (I) may be prepared by one or more of three methods:

-   -   (i) by reacting the compound of formula (I) with the desired        acid or base;    -   (ii) by removing an acid- or base-labile protecting group from a        suitable precursor of the compound of formula (I) or by        ring-opening a suitable cyclic precursor, for example, a lactone        or lactam, using the desired acid or base; or    -   (iii) by converting one salt of the compound of formula (I) to        another by reaction with an appropriate acid or base or by means        of a suitable ion exchange column.

All three reactions are typically carried out in solution. The resultingsalt may precipitate out and be collected by filtration or may berecovered by evaporation of the solvent. The degree of ionization in theresulting salt may vary from completely ionized to almost non-ionized.

The compounds of the invention may exist in both unsolvated and solvatedforms. The term ‘solvate’ is used herein to describe a molecular complexcomprising the compound of the invention and a stoichiometric amount ofone or more pharmaceutically acceptable solvent molecules, for example,ethanol. The term ‘hydrate’ is employed when said solvent is water.Included within the scope of the invention are complexes such asclathrates, drug-host inclusion complexes wherein, in contrast to theaforementioned solvates, the drug and host are present in stoichiometricor non-stoichiometric amounts. Also included are complexes of the drugcontaining two or more organic and/or inorganic components, which may bein stoichiometric or non-stoichiometric amounts. The resulting complexesmay be ionised, partially ionized, or non-ionized. For a review of suchcomplexes, see J Pharm Sci, 64 (8), 1269-1288 by Haleblian (August1975).

Hereinafter all references to compounds of formula (I) includereferences to salts, solvates and complexes thereof and to solvates andcomplexes of salts thereof.

The compounds of the invention include compounds of formula (I) ashereinbefore defined, including all polymorphs and crystal habitsthereof, prodrugs and isomers thereof (including optical, geometric andtautomeric isomers) as hereinafter defined and isotopically-labeledcompounds of formula (I).

As indicated, so-called ‘pro-drugs’ of the compounds of formula (I) arealso within the scope of the invention. Thus certain derivatives ofcompounds of formula (I) which may have little or no pharmacologicalactivity themselves can, when administered into or onto the body, beconverted into compounds of formula (I) having the desired activity, forexample, by hydrolytic cleavage. Such derivatives are referred to as‘prodrugs’. Further information on the use of prodrugs may be found in‘Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T.Higuchi and W. Stella) and ‘Bioreversible Carriers in Drug Design’,Pergamon Press, 1987 (ed. E. B Roche, American PharmaceuticalAssociation).

Prodrugs in accordance with the invention can, for example, be producedby replacing appropriate functionalities present in the compounds offormula (I) with certain moieties known to those skilled in the art as‘pro-moieties’ as described, for example, in “Design of Prodrugs” by H.Bundgaard (Elsevier, 1985).

Some examples of prodrugs in accordance with the invention include:

(i) where the compound of formula (I) contains a carboxylic acidfunctionality (—COOH), an ester thereof, for example, a compound whereinthe hydrogen of the carboxylic acid functionality of the compound offormula (I) is replaced by (C₁-C₈)alkyl;

(ii) where the compound of formula (I) contains an alcohol functionality(—OH), an ether thereof, for example, a compound wherein the hydrogen ofthe alcohol functionality of the compound of formula (I) is replaced by(C₁-C₆)alkanoyloxymethyl; and

(iii) where the compound of formula (I) contains a primary or secondaryamino functionality, an amide thereof, for example, a compound wherein,as the case may be, one or both hydrogens of the amino functionality ofthe compound of formula (I) is/are replaced by (C₁-C₁₀)alkanoyl.

Further examples of replacement groups in accordance with the foregoingexamples and examples of other prodrug types may be found in theaforementioned references. Moreover, certain compounds of formula (I)may themselves act as prodrugs of other compounds of formula (I).

Also included within the scope of the invention are metabolites ofcompounds of formula (I), that is, compounds formed in vivo uponadministration of the drug. Some examples of metabolites in accordancewith the invention include

-   -   (i) where the compound of formula (I) contains a methyl group,        an hydroxymethyl derivative thereof;    -   (ii) where the compound of formula (I) contains a tertiary amino        group, a secondary amino derivative thereof;    -   (iii) where the compound of formula (I) contains a secondary        amino group, a primary derivative thereof;    -   (iv) where the compound of formula (I) contains a phenyl moiety,        a phenol derivative thereof.

Compounds of formula (I) containing one or more asymmetric carbon atomscan exist as two or more stereoisomers. Included within the scope of thepresent invention are all stereoisomers, geometric isomers andtautomeric forms of the compounds of formula (I), including compoundsexhibiting more than one type of isomerism, and mixtures of one or morethereof. Also included is acid addition or base salts wherein thecounter ion is optically active, for example, d-lactate or 1-lysine, orracemic, for example, dl-tartrate or dl-arginine. Conventionaltechniques for the preparation/isolation of individual enantiomersinclude chiral synthesis from a suitable optically pure precursor orresolution of the racemate (or the racemate of a salt or derivative)using, for example, chiral high-pressure liquid chromatography (HPLC).

Alternatively, the racemate (or a racemic precursor) may be reacted witha suitable optically active compound, for example, an alcohol, or, inthe case where the compound of formula (I) contains an acidic or basicmoiety, an acid or base such as tartaric acid or 1-phenylethylamine. Theresulting diastereomeric mixture may be separated by chromatographyand/or fractional crystallization and one or both of thediastereoisomers converted to the corresponding pure enantiomer(s) bymeans well known to a skilled person.

Chiral compounds of the invention (and chiral precursors thereof) may beobtained in enantiomerically-enriched form using chromatography,typically HPLC (chiral columns), on an asymmetric resin with a mobilephase consisting of a hydrocarbon, typically heptane or hexane,containing from 0 to 50% by volume of isopropanol, typically from 2% to20%, and from 0 to 5% by volume of an alkylamine, typically 0.1%diethylamine. For reverse HPLC CH₃CN and H₂O, MeOH or iPrOH and H₂O areused as solvents. Concentration of the eluate affords the enrichedmixture.

Stereoisomeric conglomerates may be separated by conventional techniquesknown to those skilled in the art-see, for example, “Stereochemistry ofOrganic Compounds” by E. L. Eliel (Wiley, New York, 1994). “ChiralSeparation Techniques”. by G. Subramanian. John Wiley & Sons, 2008.“Preparative Enantioselective Chromatography” by G. B. Cox. Wiley, 2005.

Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O.

The compounds of formula (I), their pharmaceutically acceptable saltsand/or derived forms, are valuable pharmaceutically active compounds,which are suitable for the therapy and prophylaxis of various cancers,in particular melanoma, breast, prostate and colon. Compounds of theinvention may be administered as crystalline or amorphous products. Theymay be obtained, for example, as solid plugs, powders, or films bymethods such as precipitation, crystallization, freeze-drying, spraydrying, or evaporative drying. Microwave or radio frequency drying maybe used for this purpose. They may be administered alone or incombination with one or more other compounds of the invention or incombination with one or more other drugs (or as any combinationthereof). Generally, they will be administered as a formulation inassociation with one or more pharmaceutically acceptable excipients. Theterm “excipient” is used herein to describe any ingredient other thanthe compound(s) of the invention. The choice of excipient will to alarge extent depend on factors such as the particular mode ofadministration, the effect of the excipient on solubility and stability,and the nature of the dosage form. Pharmaceutical compositions suitablefor the delivery of compounds of the present invention and methods fortheir preparation will be readily apparent to those skilled in the art.Such compositions and methods for their preparation may be found, forexample, in ‘Remington's Pharmaceutical Sciences’, 19th Edition (MackPublishing Company, 1995).

Another aspect of the invention is thus a pharmaceutical compositioncomprising a compound as defined above and a pharmaceutically acceptablecarrier.

The compounds of the invention may be administered by any suitableroute.

Thus, a compound of the invention may be formulated as a pharmaceuticalcomposition for oral, buccal, intranasal, parenteral (e.g. intravenous,intramuscular or subcutaneous), topical, or rectal administration or ina form suitable for administration by inhalation or insufflation. Fororal administration, the pharmaceutical composition may take the formof, for example, a tablet or capsule prepared by conventional means witha pharmaceutically acceptable excipient such as a binding agent (e.g.,pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose); filler (e.g., lactose, microcrystalline cellulose orcalcium phosphate); lubricant (e.g., magnesium stearate, talc orsilica); disintegrant (e.g., potato starch or sodium starch glycolate);or wetting agent (e.g., sodium lauryl sulphate). The tablets may becoated by methods well known in the art. Liquid preparations for oraladministration may take the form of a, for example, solution, syrup orsuspension, or they may be presented as a dry product for constitutionwith water or other suitable vehicle before use.

Such liquid preparations may be prepared by conventional means with apharmaceutically acceptable additive such as a suspending agent (e.g.,sorbitol syrup, methyl cellulose or hydrogenated edible fats);emulsifying agent (e.g., lecithin or acacia); non-aqueous vehicle (e.g.,almond oil, oily esters or ethyl alcohol); and preservative (e.g.,methyl or propyl p-hydroxybenzoates or sorbic acid).

For buccal administration, the composition may take the form of tabletsor lozenges formulated in conventional manner. A compound of the presentinvention may also be formulated for sustained delivery according tomethods well known to those of ordinary skill in the art.

Examples of such formulations can be found in U.S. Pat. Nos. 3,538,214,4,060,598, 4,173,626, 3,119,742, and 3,492,397, which are hereinincorporated by reference in their entirety.

A compound of the invention may be formulated for parenteraladministration by injection, including using conventionalcatheterization techniques or infusion. Formulations for injection maybe presented in unit dosage form, e.g., in ampules or in multi-dosecontainers, with an added preservative. The compositions may take suchforms as suspensions, solutions or emulsions in oily or aqueousvehicles, and may contain a formulating agent such as a suspending,stabilizing and/or dispersing agent. Alternatively, the activeingredient may be in powder form for reconstitution with a suitablevehicle, e.g., sterile pyrogen-free water, before use parenteralformulations are typically aqueous solutions which may containexcipients such as salts, carbohydrates and buffering agents (preferablyto a pH of from 3 to 9), but, for some applications, they may be moresuitably formulated as a sterile non-aqueous solution or as a dried formto be used in conjunction with a suitable vehicle such as sterile,pyrogen-free water.

Inasmuch as it may desirable to administer a combination of activecompounds, for example, for the purpose of treating a particular diseaseor condition, it is within the scope of the present invention that twoor more pharmaceutical compositions, at least one of which contains acompound in accordance with the invention, may conveniently be combinedin the form of a kit suitable for coadministration of the compositions.Thus, the kit of the invention comprises two or more separatepharmaceutical compositions, at least one of which contains a compoundof formula (I) in accordance with the invention, and means forseparately retaining said compositions, such as a container, dividedbottle, or divided foil packet. An example of such a kit is the familiarblister pack used for the packaging of tablets, capsules and the like.

The kit of the invention is particularly suitable for administeringdifferent dosage forms, for example parenteral, for administering theseparate compositions at different dosage intervals, or for titratingthe separate compositions against one another. To assist compliance, thekit typically comprises directions for administration and may beprovided with a so-called memory aid.

For administration to human patients, the total daily dose of thecompounds of the invention is typically in the range 0.001 mg to 5000 mgdepending, of course, on the mode of administration. For example, anintravenous daily dose may only require from 0.001 mg to 40 mg. Thetotal daily dose may be administered in single or divided doses and may,at the physician's discretion, fall outside of the typical range givenherein.

These dosages are based on an average human subject having a weight ofabout 65 kg to 70 kg. The physician will readily be able to determinedoses for subjects whose weight falls outside this range, such asinfants and the elderly.

For the avoidance of doubt, references herein to “treatment” includereferences to curative, palliative and prophylactic treatment.

According to another embodiment of the present invention, the compoundsof the formula (I), or pharmaceutically acceptable salts, derived formsor compositions thereof, can also be used as a combination with one ormore additional therapeutic agents to be co-administered to a patient toobtain some particularly desired therapeutic end result such as thetreatment of cancers namely melanoma, breast, prostate and colon cancer.

The second and more additional therapeutic agents may also be compoundsof the formula (I), or a pharmaceutically acceptable salt, derived formsor compositions thereof, or one or more compounds known in the art forthe treatment of the conditions listed above. More typically, the secondand more therapeutic agents will be selected from a different class oftherapeutic agents.

As used herein, the terms “co-administration”, “co-administered” and “incombination with”, referring to the compounds of formula (I) and one ormore other therapeutic agents, is intended to mean, and does refer toand include the following: simultaneous administration of suchcombination of compound(s) of formula (I) and therapeutic agent(s) to apatient in need of treatment, when such components are formulatedtogether into a single dosage form which releases said components atsubstantially the same time to said patient, substantially simultaneousadministration of such combination of compound(s) of formula (I) andtherapeutic agent(s) to a patient in need of treatment, when suchcomponents are formulated apart from each other into separate dosageforms which are taken at substantially the same time by said patient,whereupon said components are released at substantially the same time tosaid patient, sequential administration of such combination compound(s)of formula (I) and therapeutic agent(s) to a patient in need oftreatment, when such components are formulated apart from each otherinto separate dosage forms which are taken at consecutive times by saidpatient with a significant time interval between each administration,whereupon said components are released at substantially different timesto said patient; and sequential administration of such combination ofcompound(s) of formula (I) and therapeutic agent(s) to a patient in needof treatment, when such components are formulated together into a singledosage form which releases said components in a controlled mannerwhereupon they are concurrently, consecutively, and/or administered atthe same and/or different times by said patient, where each part may beadministered by either the same or different route. Suitable examples ofother therapeutic agents which may be used in combination with thecompound(s) of formula (I), or pharmaceutically acceptable salts,derived forms or compositions thereof, include, but are by no meanslimited to:

-   -   Anti cancer agents used for the therapy of cancers such as        dacarbazine,    -   Nitrosourea alkylating agents, such as fotemustine    -   BRAF inhibitors such as vemurafenib or dabrafenib,    -   MEK inhibitors such as trametinib,    -   Anti-CTLA4 antibodies, namely ipilimumab

It is to be appreciated that all references herein to treatment includecurative, palliative and prophylactic treatment. The description, whichfollows, concerns the therapeutic applications to which the compounds offormula (I) may be put. A still further aspect of the present inventionalso relates to the use of the compounds of formula (I), orpharmaceutically acceptable salts, derived forms or compositionsthereof, for the manufacture of a drug having an anticancer activity. Inparticular, the present inventions concerns the use of the compounds offormula (I), or pharmaceutically acceptable salts, derived forms orcompositions thereof, for the manufacture of a drug for the treatment ofmelanoma. As a consequence, the present invention provides aparticularly interesting method to treat a mammal, including a humanbeing, with an effective amount of a compound of formula (I) or apharmaceutically acceptable salt, derived form or composition thereof.More precisely, the present invention provides a particularlyinteresting method for the treatment of a cancer disease in a mammal,including a human being, in particular the diseases and/or conditionslisted above, comprising administering said mammal with an effectiveamount of a compound of formula (I), its pharmaceutically acceptablesalts and/or derived forms. The following examples illustrate thepreparation of the compounds of the formula (I) and theirpharmacological properties

FIGURES

FIG. 1: The effect of compounds Ia-7, IIIa-3, IIIa-1, IIIa-2, IIa-2,IIa-9, Ia-4, IIa-11, IIa-12, IIa-6, compared to Ia-1, on cell viabilityon A375 melanoma cells.

FIG. 2a : Effect of compounds IIa-2, IIIa-1 and IIIa-3 on cell viabilityon melanoma cells. PLX4032 was used as a positive control.

FIG. 2b : Effect of compounds IIa-2, IIIa-1 and IIIa-3 on cell viabilityon CML cells. Imatinib was used as a positive control.

FIG. 2c : Effect of compounds IIa-2, IIIa-1 and IIIa-3 on cell viabilityon pancreatic cancer cells. Gemcitabine was used as a positive control.

FIG. 3: Effect of compound IIa-2 on the viability of KHN and FHN celllines.

FIG. 4: Western Blots showing the mechanism of death of compound IIa-2(apoptosis versus autophagy).

EXAMPLES Chemical Synthesis and Characterization

¹H and ¹³C NMR spectra were recorded on 200 or 500 Bruker AdvanceSpectrometers (200 or 500 MHz for ¹H, 50 for ¹³C). Chemical shifts areexpressed as parts per million from tetramethylsilane. Splittingpatterns have been designated as follows: s (singlet), d (doublet), t(triplet), m (multiplet) and br (broad). Coupling constants (J values)are listed in hertz (Hz). Analytical thin-layer chromatography (TLC) wasconducted on Merck (VWR) precoated silica gel 60F254 plates andcompounds were visualized with ninhydrin test and/or under ultravioletlight (254 nm). Column chromatographies were carried out on silica gel(Merck, 40-63 μm). Electrospray ionization spectrometry (ESI-MS) inpositive mode was performed on a Burker Daltonics (Esquire 3000 plus)apparatus. HPLC analyses were recorded on waters instruments usingcolumns with different sizes.

HPLC Methods:

Solvent A: H₂O (0.1% formic acid) and solvent B: CH₃CN (0.1% formicacid).Method 1: 0% B to 100% B over 10 min, 100% B for 5 min then from 100% Bto 0% B over 1 min (16 min in total).Method 2: 25% B to 100% B over 13 min, 100% B for 5 min then from 100% Bto 0% B over 1 min (19 min in total).Method 3: 25% B for 3 min then 25% B to 95% B over 5 min, 100% B for 5min then from 100% B to 0% B over 1 min (14 min in total).Method 4: 25% B to 100% B over 14 min, 100% B for 5 min then from 100% Bto 0% B over 1 min (19 min in total).General Procedure for the Formation of Sulfonamides (I) To a solution ofN-(4-(3-aminophenyl)thiazol-2-yl)acetamide (1 eq.) under argon inanhydrous DMF (0.1M) were added triethylamine (1.6 eq.) and thecorresponding sulfonyl chloride (1.2 eq). The reaction mixture wasallowed to react at r.t. until complete conversion of the startingmaterial (from 2 h to 48 h). DMF was removed under reduced pressure andthe crude material was purified by silica gel flash chromatography toafford the pure corresponding sulfonamides.

General Procedure for the Sonogashira Coupling (II)

To a suspension of halogenated derivative (Ia-3 or Ia-4) (1 eq.) in amixture of Et₃N/benzene or toluene (1/1, [0.17M]) under argon were addedPd(PPh₃)₄ (15% mol.), Copper (I) iodide (15% mol.) and correspondingalkyne (5 eq.). The resulting mixture was stirred at 80° C. untilcomplete conversion of the starting material (2 h approx.). The reactionmixture was then cooled to r.t. and all volatiles were removed underreduced pressure and the crude material was purified by silica gel flashchromatography to afford the pure corresponding coupling product.

Synthetic Procedures and Characterizations:

Product reference Structure JM010 (2a-1)

2-bromo-1-(3-nitrophenyl)ethanone (2a-1). To a suspension ofcommercially available 3-nitroacetophenone (4.82 g, 29.19 mmol) inanhydrous diethyl ether (25 mL) was added aluminum chloride (0.16 g,1.20 mmol). The reaction mixture was then cooled to 0° C. beforedropwise addition of bromine (1.50 mL, 29.19 mmol). The reaction wasstirred for 1 h at room temperature until complete conversion of thestarting material (′H NMR monitoring) and quenched quickly to avoiddi-brominated compound formation. After addition of water (30 mL), themixture was extracted with diethyl ether (3×30 mL) and dried with MgSO₄to afford compound 2a-1 as a yellowish brown solid (5.73 g, 97% yield).¹H NMR (CDCl₃, 200 MHz): δ 4.50 (s, 2H, H₁), 7.73 (t, J=8.0 Hz, 1H andH₇), 8.31 (ddd, J=7.8, 1.7, 1.1 Hz, 1H, H₈), 8.43 (ddd, J=8.2, 2.3, 1.1Hz, 1H, H₆), 8.76 (t, J=1.9 Hz, 1H, H₄). ¹³C NMR (CDCl₃, 200 MHz): δ30.4, 123.9, 128.2, 130.4, 134.6, 135.2, 148.6, 189.5. MS-ESI (m/z):[M+H]⁺=244.9.

Product reference Structure SBN (4a-1)

N-(4-(3-nitrophenyl)thiazol-2-yl)acetamide (4a-1). To a solution of2-bromo-1-(3-nitrophenyl)ethanone (4.27 g, 17.50 mmol) in ethanol(technical grade, 150 mL) was added N-acethylthiourea (2.07 g, 17.50mmol). The reaction mixture was heated to 80° C. for 30 minutes thenallowed to cool down to room temperature. The precipitate was filteredand washed with an ice-cooled solution of 1:1 ethanol/diethyl-ether (200mL) affording compound 4a-1 as a yellow solid (4.24 g, 92% yield). ¹HNMR (DMSO-d₆, 200 MHz): δ 2.17 (s, 3H, H₁), 7.72 (t, J=8.2 Hz, 1H, H₁₀),7.92 (s, 1H, H₄), 8.16 (dd, J=8.1, 2.4 Hz, 1H, H₁₁), 8.33 (d, J=8.1 Hz,1H, H₉), 8.71 (d, J=2.1 Hz, 1H, H₇), 12.38 (s, 1H, NH_(acetyl)). ¹³C NMR(DMSO-d₆, 50 MHz): δ 22.5, 110.4, 120.0, 122.3, 130.4, 131.7, 135.8,146.3, 148.3, 158.4, 168.8. HRMS-ESI (m/z): [M+H]⁺ Calcd forC₁₁H₁₀O₃N₃S, 264.0437; found: 264.0439.

Product reference Structure KM085 (4a-2)

N-methyl-4-(3-nitrophenyl)thiazol-2-amine (4a-2). To a solution of2-bromo-1-(3-nitrophenyl)ethanone (4.0 g, 16.4 mmol) in ethanol(technical grade, 150 mL) was added N-methylthiourea (1.7 g, 19.7 mmol).The reaction mixture was stirred for 2 h at r.t. and ethanol was removedunder reduced pressure. The precipitate was filtered and washed with anice-cooled (4° C.) solution of 1:1 ethanol/diethyl-ether (200 mL)affording compound 4a-2 as an orange solid (3.9 g, quant.) pure enoughto carry on the synthesis. ¹H NMR (CD₃OD, 200 MHz): δ 3.21 (s, 3H, H₁),7.38 (s, 1H, H₃), 7.78 (t, J=8.1 Hz, 1H, H₉), 8.23-8.05 (m, 1H, H₁₀),8.34 (dd, J=8.3, 2.3 Hz, 1H, H₈), 8.60 (t, J=2.1 Hz, 1H, H₆). ¹³C NMR(CD₃OD, 50 MHz): δ 33.3, 106.3, 122.5, 125.6, 131.4, 131.9, 133.8,139.3, 150.1, 172.6. MS-ESI (m/z): [M+H]⁺=235.9.

Product reference Structure KM081 (4a-3)

4-(3-nitrophenyl)-N-phenylthiazol-2-amine (4a-3). To a solution of2-bromo-1-(3-nitrophenyl)ethanone (4.30 g, 17.70 mmol) in ethanol(technical grade, 150 mL) was added N-phenylthiourea (2.69 g, 17.70mmol). The reaction mixture was stirred for 2 h at r.t. and ethanol wasremoved under reduced pressure. The precipitate was filtered and washedwith an ice-cooled (4° C.) solution of 1:1 ethanol/diethyl-etheraffording compound 4a-3 as a yellow solid (5.22 g, quant.). ¹H NMR(DMSO-d₆, 200 MHz): δ 7.80 (t, J=7.3 Hz, 1H, H, 2), 8.17 (t, J=8.3 Hz,2H, H₂), 8.64-8.43 (m, 4H, H₁, H₃ and H₆), 8.97 (ddd, J=8.2, 2.4, 1.0Hz, 1H, H₁₃), 9.18 (dt, J=7.8, 1.3 Hz, 1H, H₁₁), 9.51 (t, J=2.0 Hz, 1H,H₉), 11.22 (s, 1H, NH_(thiazole)). ¹³C NMR (DMSO-d₆, 50 MHz): δ 105.7,117.0 (2C), 119.9, 121.5, 122.1, 129.0 (2C), 130.3, 131.8, 136.0, 141.0,147.6, 148.3, 163.6. MS-ESI (m/z): [M+H]⁺=298.0; [M+Na]⁺=320.0.

Product name Structure DRM46- piper (5a-4)

N-(4-(3-nitro-4-(4-methylpiperazin-1-yl)phenyl)thiazol-2-yl)acetamide(4a-4). To a yellow suspension ofN-(4-(3-nitro-4-fluorophenyl)thiazol-2-yl)acetamide (336 mg, 1.20 mmol)in DMSO (25 mL) was added N-methylpiperazine (167 μL, 1.32 mmol). Themixture was sonicated at r.t. in the ultrasonic bath for 20 min, afterwhich it became a limpid orange solution. Cold basic water (made with160 mL water+40 mL saturated aq. Na₂CO₃) was added to the mixture andthe product was extracted with ethyl acetate three times. The combinedorganic layers were dried with MgSO₄, and concentrated under reducedpressure to afford an orange solid corresponding to substitution product4a-4 (412 mg, 95%). ¹H NMR (DMSO-d₆, 200 MHz): δ 2.16 (s, 3H, H₁),3.11-2.94 (m, 4H, H₁₃), 3.33 (s, 3H, H, 4), 3.79-3.59 (m, 4H, H₁₂), 7.38(d, J=8.7 Hz, 1H, H₁₀), 7.69 (s, 1H, H₄), 8.09 (dd, J=8.6, 2.2 Hz, 1H,H₁₁), 8.34 (d, J=2.1 Hz, 1H, H₇), 12.29 (s, 1H, NH_(thiazole)). ¹³C NMR(DMSO-d₆, 50 MHz): δ 22.58, 40.8, 51.3 (2C), 66.0, 108.2, 121.6, 122.5,128.0, 130.6, 124.2, 144.4, 146.4, 158.2, 168.7. MS-ESI (m/z):[M+H]⁺=362.5.

Product name Structure DRM46- morpho (4a-5)

N-(4-(3-nitro-4-morpholinophenyl)thiazol-2-yl)acetamide (4a-5). To ayellow suspension of N-(4-(3-nitro-4-fluorophenyl)thiazol-2-yl)acetamide(336 mg, 1.20 mmol) in DMSO (25 mL) was added morpholine (114 μL, 1.32mmol). The mixture was sonicated at r.t. in the ultrasonic bath for 30min, after which it became a limpid orange solution. Cold acidic water(200 mL; acidified to pH 3 using diluted aq. HCl) were added to themixture. The orange precipitate that formed was filtered and washed withwater. The solid obtained was dried at 50° C. overnight to afford puresubstitution product 4a-5 (401 mg, 96%). ¹H NMR (DMSO-d₆, 200 MHz): δ2.16 (s, 3H, H₁), 3.03 (t, J=4.6 Hz, 4H, H₁₃), 3.71 (t, J=4.5 Hz, 4H,H₁₂), 7.38 (d, J=8.7 Hz, 1H, H₁₀), 7.69 (s, 1H, H₄), 8.09 (dd, J=8.6,2.2 Hz, 1H, KO, 8.34 (d, J=2.1 Hz, 1H, H₇), 12.29 (s, 1H,NH_(thiazole)). ¹³C NMR (DMSO-d₆, 50 MHz): δ 22.5, 51.3 (2C), 66.0 (2C),108.2, 121.6, 122.6, 128.1, 130.6, 142.2, 144.4, 146.4, 158.2, 168.7.MS-ESI (m/z): [M+H]⁺=349.5.

Product reference Structure SBA (5a-1)

N-(4-(3-aminophenyl)thiazol-2-yl)acetamide (5a-1). To a suspension of4a-1 (2.00 g, 7.60 mmol) and 10% Pd/C (0.20 g, 10% wt.) in methanol(technical grade, 76 mL) under stirring at 0° C., was added carefully byportions sodium borohydride (1.44 g, 38.00 mmol). The reaction mixturewas stirred at 0° C. until complete dissolution of sodium borohydrideand was then allowed to react 3 h at r.t. After completion of thereaction monitored by TLC (CH₂Cl₂/MeOH 90:10), the mixture was filteredthrough a pad of Celite®, concentrated under reduced pressure andpurified by silica gel flash chromatography eluted with CH₂Cl₂/MeOH(99:1 to 95:5) to afford 5a-1 as a yellow-white solid (1.47 g, 83%). ¹HNMR (DMSO-d₆, 200 MHz): δ 2.16 (s, 3H, H₁), 5.13 (s, 2H, NH₂), 6.52 (dt,J=6.4, 2.5 Hz, H₉), 7.16-6.94 (m, 3H, H₇, H₁₀ and H₁₁), 7.35 (s, 1H,H₄), 12.22 (s, 1H, NH_(acetyl)). ¹³C NMR (DMSO-d₆, 50 MHz): δ 22.5,107.0, 111.3, 113.6, 113.6, 127.2, 134.9, 148.9, 149.6, 157.6, 168.6.HRMS-ESI (m/z): [M+H]⁺ Calcd for C₁₁H₁₂N₃OS, 234.0696; found: 234.0700.

Product name Structure KM086

N-methyl-4-(3-aminophenyl)thiazol-2-amine (5a-2). To a suspension of4a-2 (4.5 g, 19.1 mmol) and 10% Pd/C (0.45 g, 10% wt.) in methanol(technical grade, 191 mL) under stirring at 0° C., was added carefullyby portions sodium borohydride (3.62 g, 95.5 mmol). The reaction mixturewas stirred at 0° C. until complete dissolution of sodium borohydrideand was then allowed to react 3 h at r.t. After completion of thereaction monitored by TLC (CH₂Cl₂/MeOH 90:10), the mixture was filteredthrough a pad of Celite®, concentrated under reduced pressure andpurified by silica gel flash chromatography eluted with CH₂Cl₂/MeOH(99:1 to 95:5) to afford 5a-2 as an orange solid (1.75 g, 45%). ¹H NMR(DMSO-d₆, 200 MHz): δ 2.85 (d, J=4.8 Hz, 3H, H₁), 5.06 (s, 2H, NH₂),6.53-6.38 (m, 1H, H₈), 6.83 (s, 1H, H₃), 7.01-6.94 (m, 2H, H₉ and H₁₀),7.09-7.03 (m, 1H, H₆). ¹³C NMR (DMSO-d₆, 50 MHz): δ 31.0, 99.9, 111.5,113.1, 113.6, 128.8, 135.5, 148.6, 151.1, 169.0.

Product reference Structure AM360

4-(3-aminophenyl)-N-phenylthiazol-2-amine (5a-3). To a suspension of4a-3 (500.0 mg, 1.68 mmol) and 10% Pd/C (50.0 mg, 10% wt.) in methanol(technical grade, 35 mL) under stirring at 0° C., was added carefully byportions sodium borohydride (445.7 mg, 11.76 mmol). The reaction mixturewas stirred at 0° C. until complete dissolution of sodium borohydrideand was then allowed to react 3 h at r.t. After completion of thereaction monitored by TLC (CH₂Cl₂/MeOH 90:10), the mixture was filteredthrough a pad of Celite®, concentrated under reduced pressure andpurified by silica gel flash chromatography eluted with CH₂Cl₂/MeOH(99:1 to 95:5) to afford 5a-3 as a pale yellow solid (200.5 mg, 45%). ¹HNMR (CDCl₃, 200 MHz): δ 3.30 (s, 2H, NH₂), 6.46 (ddd, J=7.6, 2.3, 1.3Hz, 1H, H₁₁), 6.58 (s, 1H, H₆), 7.23-6.82 (m, 8H, H₁, H₂, H₃, H₉, H₁₂and H₁₃), 8.51 (s, 1H, NH_(thiazole)). ¹³C NMR (CDCl₃, 50 MHz): δ 101.8,113.2, 114.9, 116.7, 118.4 (2C), 122.9, 129.4 (2C), 129.6, 135.6, 140.6,146.7, 151.3, 165.2. MS-ESI (m/z): [M+H]⁺=267.9.

Product name Structure AM370 (5a-4)

N-(4-(3-amino-4-(4-methylpiperazin-1-yl)phenyl)thiazol-2-yl)acetamide(5a-4). To a suspension of 4a-4 (300.0 mg, 0.83 mmol) and 10% Pd/C (30.0mg, 10% wt.) in methanol (technical grade, 17 mL) under stirring at 0°C., was added carefully by portions sodium borohydride (156.9 mg, 4.15mmol). The reaction mixture was stirred at 0° C. until completedissolution of sodium borohydride and was then allowed to react 3 h atr.t. After completion of the reaction monitored by TLC (CH₂Cl₂/MeOH90:10), the mixture was filtered through a pad of Celite®, concentratedunder reduced pressure and purified by silica gel flash chromatographyeluted with CH₂Cl₂/MeOH (3% Et₃N, 95:5 to 90:10) to afford 5a-4 as abrown powder (120.0 mg, 44%). ¹H NMR (CD₃OD, 200 MHz): δ 2.20 (s, 3H,H₁), 2.44 (s, 3H, H₁₄), 2.76 (s, 4H, H₁₃), 2.97 (t, J=4.9 Hz, 4H, H₁₂),6.99 (d, J=8.2 Hz, 1H, H₁₀), 7.15 (s, 1H, H₄), 7.22 (dd, J=8.2, 2.0 Hz,1H, 7.30 (d, J=2.0 Hz, 1H, H₇). ¹³C NMR (CD₃OD, 50 MHz): δ 22.8, 45.9,51.4 (2C), 56.5 (2C), 107.6, 114.4, 117.7, 120.9, 132.8, 140.0, 143.3,151.5, 159.3, 171.0. MS-ESI (m/z): [M+H]⁺=332.0; [M+Na]⁺=354.0.

Product name Structure CR068 (5a-5)

N-(4-(3-amino-4-morpholinophenyl)thiazol-2-yl)acetamide (5a-5). To ayellow suspension of nitroaryl derivative 4a-5 (200 mg, 0.574 mmol) inmethanol (technical grade, 20 mL) was added Pd/C (30 mg, 10% wt.), thensodium borohydride (130 mg, 3.44 mmol). The mixture was stirred at r.t.for 1 h after which it became a dark green suspension. The mixture wasfiltered through Celite® and the filtrate was evaporated. The residuewas partitioned between diluted aq. Na₂CO₃ and ethyl acetate. Theaqueous phase was extracted with ethyl acetate twice. The combinedorganic layers were dried with MgSO₄, and concentrated under reducedpressure. Purification by flash chromatography (CH₂Cl₂:CH₃OH, 100:0 to94:6) afforded the pure desired aniline 5a-5 as a pale yellow solid(165.5 mg, 91%). ¹H NMR (DMSO-d₆, 200 MHz): δ 2.16 (s, 3H, H₁),2.99-2.69 (m, 4H, H₁₂), 3.74 (dd, J=5.8, 3.1 Hz, 4H, H₁₃), 4.85 (s, 2H,NH₂), 6.89 (d, J=8.2 Hz, 1H, H₁₀), 7.09 (dd, J=8.1, 2.0 Hz, 1H, H₁₁),7.22 (d, J=6.5 Hz, 2H, H₄ and H₇), 12.21 (s, 1H, NH_(thiazole)). ¹³C NMR(DMSO d, 50 MHz): δ 22.5, 50.9 (2C), 66.6 (2C), 106.0, 111.9, 114.6,119.1, 130.3, 137.9, 142.2, 149.5, 157.6, 168.5. MS-ESI (m/z):[M+H]⁺=319.0; [M+Na]⁺=341.0.

Product ref Structure JG28- F4 (Ia-1)

N-(4-(3-(phenylsulfonamido)phenyl)thiazol-2-yl)acetamide (Ia-1). Thegeneral procedure for the formation of sulfonamides (I) was followedusing 5a-1 (102 mg, 0.44 mmol), benzenesulfonyl chloride (67.6 μL, 0.53mmol) as reagents and triethylamine. Purification by silica gel flashchromatography (CH₂Cl₂:EtOAc; 100:0 to 80:20) afforded the titledcompound as a white fine powder (28.8 mg, 37%). ¹H NMR (Acetone-d₆, 200MHz): δ 2.28 (s, 3H, H₁), 7.10 (ddd, J=8.0, 2.2, 1.1 Hz, 1H, H₉),7.30-7.20 (m, 1H, H₁₀), 7.37 (s, 1H, H₄), 7.64-7.46 (m, 4H, H₁₁, H₁₄ andH₁₅), 7.90-7.79 (m, 3H, H₇ and H₁₃), 9.12 (s, 1H, NH_(sulfonamide)),11.21 (s, 1H, NH_(acetyl)). ¹³C NMR (Acetone-d₆, 50 MHz): δ 22.5, 108.5,117.6, 119.7, 121.6, 126.7 (2C), 129.3 (2C), 129.5, 132.9, 135.3, 138.2,139.5, 148.1, 158.0, 168.7. HRMS-ESI (m/z): [M+H]⁺ Calcd forC₁₇H₁₆N₃O₃S₂, 374.0628; Found: 374.0627.

Product ref Structure JG25 (Ia-2)

N-(4-(3-((4-methylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(Ia-2). The general procedure for the formation of sulfonamides (I) wasfollowed using 5a-1 (102 mg, 0.44 mmol), tosyl chloride (101.0 mg, 0.53mmol) as reagents and triethylamine. Purification by silica gel flashchromatography (CH₂Cl₂:EtOAc; 100:0 to 80:20) afforded the titledcompound as a white powder (122.7 mg, 72%). ¹H NMR (Acetone-d₆, 200MHz): δ 2.28 (s, 3H, H₁), 2.33 (s, 3H, H₁₆), 7.10 (ddd, J=8.0, 2.2, 1.1Hz, 1H, H₉), 7.34-7.20 (m, 3H, H₁₀ and H₁₄), 7.37 (s, 1H, H₄), 7.59(ddd, J=7.7, 1.6, 1.1 Hz, 1H, 7.75-7.66 (m, 2H, H₁₃), 7.87 (ddd, J=2.2,1.6, 0.5 Hz, 1H, H₇), 9.04 (s, 1H, NH_(sulfonamide)), 11.17 (s, 1H,NH_(acetyl)). ¹³C NMR (Acetone-d₆, 50 MHz): 20.9, 22.5, 108.4, 117.4,119.5, 121.5, 126.7 (2C), 129.5, 129.7 (2C), 135.2, 136.6, 138.3, 143.3,148.2, 158.1, 168.8. HRMS-ESI (m/z): [M+H]⁺ Calcd for C₁₈H₁₈N₃O₃S₂,388.0784; Found: 388.0788.

Product ref Structure CRO51 (Ia-3)

N-(4-(3-((4-iodophenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide (Ia-3).The general procedure for the formation of sulfonamides (I) was followedusing 5a-1 (1.50 g, 6.44 mmol), 4-iodobenzenesulfonyl chloride (2.33 g,7.73 mmol) as reagents and triethylamine. Purification by silica gelflash chromatography (CH₂Cl₂:EtOAc; 100:0 to 70:30) afforded the titledcompound as a white powder (1.85 g, 60%). ¹H NMR (DMSO-d₆, 200 MHz):2.17 (s, 3H, H₁), 7.05-6.96 (m, 1H, H₉), 7.27 (t, J=7.9 Hz, 1H, H₁₀),7.62-7.45 (m, 3H, H₄, H₁₁ and H₁₄), 7.69 (t, J=1.9 Hz, 1H, H₇),7.99-7.88 (m, 1H, H₁₃), 10.45 (s, 1H, NH_(sulfonamide)), 12.26 (s, 1H,NH_(acetyl)). ¹³C NMR (DMSO-d₆, 50 MHz): δ 22.5, 101.2, 108.5, 117.7,119.8, 121.8, 128.3 (2C), 129.6, 135.3, 137.9, 138.2 (2C), 139.0, 148.0,158.0, 168.7. HRMS-ESI (m/z): [M+H]⁺ Calcd for C₁₇H₁₅IN₃O₃S₂, 499.9594;Found: 499.9591.

Product ref Structure CRO48 (Ia-4)

N-(4-(3-((4-bromophenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(Ia-4). The general procedure for the formation of sulfonamides (I) wasfollowed using 5a-1 (1.00 g, 4.29 mmol), 4-bromobenzenesulfonyl chloride(1.20 g, 4.72 mmol) as reagents and triethylamine. Purification bysilica gel flash chromatography (CH₂Cl₂:EtOAc; 100:0 to 70:30) affordedthe titled compound as a white powder (1.49 g, 77%).R_(f)(EtOAc/cyclohexane, 1/1, v/v)=0.44. ¹H NMR (CD₃OD, 200 MHz): δ 2.22(s, 3H, H₁), 7.06-6.92 (m, 1H, H₉), 7.31-7.10 (m, 2H, H₁₀ and H₄),7.73-7.46 (m, 6H, H₇, H₁₁, H₁₃, H₁₄ and H₁₅). ¹³C NMR (CD₃OD, 50 MHz): δ21.9, 107.9, 118.6, 120.2, 122.3, 127.3 (2C), 128.3, 129.2 (2C), 131.9,135.4, 137.2, 138.3, 148.7, 158.0, 169.3. HRMS-ESI (m/z): [M+H]⁺ Calcdfor C₁₇H₁₁O₃N₃BrS₂, 451.9733; Found: 451.9745.

Product ref Structure AM77 (Ia-5)

N-(4-(3-((2,3-dihydro-1H-indene)-5-sulfonamido)phenyl)thiazol-2-yl)acetamide(Ia-5). The general procedure for the formation of sulfonamide (I) wasfollowed using 5a-1 (75 mg, 0.32 mmol), indane-5-sulfonyl chloride (83.0mg, 0.39 mmol) as reagents and triethylamine. Purification by silica gelflash chromatography (CH₂Cl₂:EtOAc; 100:0 to 80:20) afforded the titledcompound as a white powder (98 mg, 74%). ¹H NMR (DMSO-d₆, 200 MHz): δ1.96 (p, J=7.4 Hz, 2H, H₁₉), 2.14 (s, 3H, H₁), 2.82 (t, J=7.4 Hz, 4H,H₁₈), 7.01 (d, J=8.9 Hz, 1H, H₉), 7.38-7.17 (m, 2H, H₁₀ and H₁₄),7.76-7.41 (m, 5H, H₄, H₇, H₁₁, H₁₃ and H₁₇), 10.29 (s, 1H,NH_(sulfonamide)), 12.25 (s, 1H, NH_(acetyl)). ¹³C NMR (DMSO-d₆, 50MHz): δ 22.5, 24.9, 32.0, 108.4, 117.2, 119.2, 121.3, 122.4, 124.8,125.0, 129.5, 135.2, 135.2, 137.5, 138.4, 145.0, 148.1, 149.5, 158.0,168.7. HRMS-ESI (m/z): [M−H]⁺ Calcd for C₂₀H₂₀N₃O₃S₂, 414.0941; Found:414.0938.

Product name Structure AM69 (Ia-6)

N-(4-(3-([1,1′-biphenyl]-4-sulfonamido)phenyl)thiazol-2-yl)acetamide(Ia-6). The general procedure for the formation of sulfonamide (I) wasfollowed using 5a-1 (100 mg, 0.43 mmol), biphenyl-4-sulfonyl chloride(130.0 mg, 0.51 mmol) as reagents and triethylamine. Purification bysilica gel flash chromatography (CH₂Cl₂:EtOAc; 80:20 to 60:40) affordedthe titled compound as a white powder (85.1 mg, 43%). ¹H NMR(Acetone-d₆, 200 MHz): δ 2.28 (s, 3H, H₁), 7.19-7.10 (m, 1H, H₉), 7.28(t, J=7.8 Hz, 1H, H₁₀), 7.54-7.37 (m, 4H, H₄, H₁₉ and H₁₈), 7.73-7.58(m, 3H, H₁₁ and H₁₇), 7.94-7.76 (m, 5H, H₇, H₁₃ and H₁₄). ¹³C NMR(DMSO-d₆, 50 MHz): δ 22.5, 108.5, 117.5, 119.5, 121.6, 127.1 (2C), 127.3(2C), 127.4 (2C), 128.6, 129.1 (2C), 129.6, 135.3, 138.2, 138.2, 138.3,144.3, 148.1, 158.0, 168.7. HRMS-ESI (m/z): [M+H]⁺ Calcd forC₂₃H₂₀N₃O₃S₂, 450.0941; Found: 450.0935.

Product name Structure AM107 (Ia-7)

N-(4-(3-((4′-methyl-[1,1′-biphenyl])-4-sulfonamido)phenyl)thiazol-2-yl)acetamide(Ia-7). The general procedure for the formation of sulfonamide (I) wasfollowed using 5a-1 (102 mg, 0.44 mmol), 4′-methylbiphenyl-4-sulfonylchloride (205.0 mg, 0.77 mmol) as reagents and triethylaminePurification by silica gel flash chromatography (CH₂Cl₂:EtOAc; 100:0 to70:30) afforded the titled compound as a white powder (110.5 mg, 37%).¹H NMR (Acetone-d₆, 200 MHz): δ 2.27 (s, 3H, H₁), 2.34 (s, 3H, H₂₀),7.19-7.12 (m, 1H, H₉), 7.31-7.22 (m, 3H, H₁₀ and H₁₈), 7.38 (s, 1H, H₄),7.65-7.49 (m, 3H, H₁₁ and H₁₇), 7.81-7.71 (m, 2H, H₁₄), 7.97-7.80 (m,3H, H₇ and H₁₃). ¹³C NMR (Acetone-d₆, 50 MHz): δ 21.1, 22.9, 108.8,119.3, 121.0, 122.9, 127.9 (2C), 128.0 (2C), 128.7 (2C), 130.3, 130.6(2C), 136.8, 137.0, 139.3 (2C), 139.3, 146.0, 149.8, 159.0, 169.2.HRMS-ESI (m/z): [M−H]^(°) Calcd for C₂₄H₂₂N₃O₃S₂, 464.1097; Found:464.1093.

Product ref Structure CRO46 (Ia-8)

N-(4-(3-((3-bromophenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(Ia-8). The general procedure for the formation of sulfonamide (I) wasfollowed using 5a-1 (583 mg, 2.50 mmol), 3-bromobenzenesulfonyl chloride(405 μL, 2.80 mmol) as reagents and triethylamine. Purification bysilica gel flash chromatography (cyclohexane:EtOAc; 100:0 to 50:50) toafforded the titled compound as a white powder (991 mg, 88%). R_(f) (E tOA c/cyclo hexane, 1/1, v/v)=0.45. ¹H NMR (CDCl₃-CD₃OD (1:1), 200 MHz):δ 2.21 (s, 3H, H₁), 7.01 (ddd, J=8.0, 2.3, 1.1 Hz, 1H, H₉), 7.12 (s, 1H,H₄), 7.26 (dt, J=10.3, 7.9 Hz, 2H, H₁₀ and H₁₆), 7.72-7.43 (m, 4H, H₁₁,H₇, H₁₅ and H₁₇), 7.92 (t, J=1.9 Hz, 1H, H₁₃). ¹³C NMR (CDCl₃-CD₃OD(1:1), 50 MHz): δ 21.8, 107.9, 118.7, 120.2, 122.3, 122.4, 125.3, 129.2,129.5, 130.2, 135.4, 135.4, 137.1, 141.1, 148.6, 158.0, 169.3. HRMS-ESI(m/z): [M+H]⁺ Calcd for C₁₇H₁₅BrN₃O₂S₂, 451.9733; Found: 451.9745.

Product ref Structure AM359 (Ia-9)

4-bromo-N-(3-(2-(methylamino)thiazol-4-yl)phenyl)benzenesulfonamide(Ia-9). The general procedure for the formation of sulfonamides (I) wasfollowed using 5a-2 (500.0 mg, 2.43 mmol), 4-bromobenzenesulfonylchloride (685.5 mg, 2.68 mmol) as reagents and triethylamine.Purification by silica gel flash chromatography (CH₂Cl₂:EtOAc; 100:0 to70:30) afforded the titled compound as a white powder (376.8 mg, 37%).¹H NMR (Acetone-d₆, 200 MHz): 2.97 (s, 1H, H₁), 6.87 (s, 2H, H₃ andNH_(thiazole)), 7.13 (ddd, J=7.9, 2.0, 1.1 Hz, 1H, H₈), 7.24 (t, J=7.8Hz, 1H, H₉), 7.59 (d, J=7.7 Hz, 1H, H₁₀), 7.84-7.64 (m, 4H, H₁₂ andH₁₃), 8.00 (s, 1H, H₆), 9.35 (s, 1H, NH_(sulfonamide)). ¹³C NMR(Acetone-d₆, 50 MHz): 31.8, 102.1, 119.7, 120.7, 123.2, 127.9, 130.0(2C), 130.1, 133.2 (2C), 137.5, 138.7, 140.3, 151.4, 170.7. MS-ESI(m/z): [M+H]⁺=424.0.

Product ref Structure AM363 (Ia-10)

4-bromo-N-(3-(2-(phenylamino)thiazol-4-yl)phenyl)benzenesulfonamide(Ia-10). The general procedure for the formation of sulfonamides (I) wasfollowed using 5a-3 (151.0 mg, 0.57 mmol), 4-bromobenzenesulfonylchloride (161.0 mg, 0.63 mmol) as reagents and triethylamine.Purification by silica gel flash chromatography (CH₂Cl₂:EtOAc; 100:0 to70:30) afforded the titled compound as a white powder (135.2 mg, 49%).¹H NMR (Acetone-d₆, 200 MHz): 7.01 (t, J=7.4 Hz, 1H, Hu), 7.22-7.07 (m,2H, H₁₁ and H₆), 7.42-7.25 (m, 3H, H₁ and H₂), 7.85-7.62 (m, 7H, H₃,H₁₅, H₁₆ and H₁₃), 7.93 (t, J=1.9 Hz, 1H, H₉), 9.20 (s, 1H,NH_(sulfonamide)), 9.36 (s, 1H, NH_(thiazole)). ¹³C NMR (Acetone-d₆, 50MHz): δ 103.8 (2C), 118.3 (2C), 119.6, 120.9, 122.6, 123.1, 128.0, 129.9(2C), 130.0, 133.2 (2C), 137.1, 138.9, 140.2, 142.4, 151.3, 164.5.MS-ESI (m/z): [M+H]⁺=486.0; [M+Na]⁺=508.0

Product ref Structure AM371 (Ia-11)

N-(4-(4-(4-methylpiperazin-1-yl)-3-((4-pentylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide (Ia-11). The general procedure for the formation ofsulfonamide (I) was followed using 5a-4 (72.1 mg, 0.22 mmol),4-pentylphenylsulfonyl chloride (59.2 mg, 0.24 mmol) as reagents andtriethylamine Purification by silica gel flash chromatography(CH₂Cl₂:CH₃OH; 95:5 to 90:10) afforded the titled compound as a whitepowder (78.5 mg, 68%). ¹H NMR (CD₃OD, 200 MHz): δ 0.84 (t, J=6.8 Hz, 3H,H₂₃), 1.36-1.10 (m, 4H, H₂, and H₂₂), 1.51 (quint., J=7.5 Hz, 2H, H₂₀),2.22 (s, 3H, H₁), 2.33 (s, 3H, H₁₄), 2.67-2.41 (m, 10H, H₁₂, H₁₃ andH₁₉), 7.13 (d, J=8.3 Hz, 1H, H₁₀), 7.24 (t, J=4.1 Hz, 3H, H₄ and H₁₇),7.56 (dd, J=8.3, 2.0 Hz, 1H, H₁₁), 7.75-7.62 (m, 2H, H₁₆), 8.17 (d,J=2.0 Hz, 1H, H₇). ¹³C NMR (CD₃OD, 50 MHz): δ 14.5, 22.8, 23.6, 32.1,32.5, 36.7, 46.2, 53.2 (2C), 56.5 (2C), 108.9, 119.8, 123.4, 124.0,128.3 (2C), 130.3 (2C), 133.9, 134.6, 138.4, 144.2, 150.4, 150.5, 159.6,171.0. MS-ESI (m/z): [M+H]⁺=542.2; [M+Na]⁺=564.2.

Product name Structure CRO69 (Ia-12)

N-(4-(4-morpholino-3-((4-pentylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(Ia-12). The general procedure for the formation of sulfonamide (I) wasfollowed using 5a-5 (81.0 mg, 0.25 mmol), 4-pentylphenylsulfonylchloride (76.0 mg, 0.31 mmol) as reagents and triethylamine Purificationby silica gel flash chromatography (CH₂Cl₂:CH₃OH; 100:0 to 94:6)afforded the titled compound as a white powder (75.2 mg, 56%). ¹H NMR(CD₃OD, 200 MHz): δ 0.86 (t, J=6.9 Hz, 3H, H₂₂), 1.42-1.14 (m, 4H, H₂₀and H₂₁), 1.53 (quint, J=7.3 Hz, 2H, H₁₉), 2.22 (s, 3H, H₁), 2.45 (dd,J=5.7, 3.4 Hz, 4H, H₁₂), 2.60 (t, J=7.6 Hz, 2H, H₁₈), 3.78-3.61 (m, 4H,H₁₃), 7.16 (d, J=8.3 Hz, 1H, H₁₀), 7.27 (t, J=4.2 Hz, 3H, H4 and H₁₆),7.60 (dd, J=8.3, 2.1 Hz, 1H, H₁₁), 7.77-7.60 (m, 2H, H₁₅), 8.17 (d,J=2.0 Hz, 1H, H₇). 13C NMR (CD3OD, 50 MHz): δ 14.3, 22.6, 23.5, 32.0,32.4, 36.5, 53.9 (2C), 68.3 (2C), 108.7, 120.1, 123.3, 123.9, 128.2(2C), 130.2 (2C), 133.7, 134.5, 138.4, 144.4, 150.3, 150.3, 159.5,170.9. MS-ESI (m/z): [M+H]+=529.2; [M+Na]+=551.3.

Product ref Structure CRO24 (IIa-1)

N-(4-(3-((4-(hex-1-yn-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(Ha-1). The general procedure for Sonogashira coupling was followedusing aryl halide 1a-3 (150.0 mg, 0.33 mmol), Pd(PPh₃)₄ (57.8 mg, 15%mol.), copper (I) iodide (9.5 mg, 15% mol.) and hex-1-yne (188.0 μL,1.66 mmol). Purification by silica gel flash chromatography(CH₂Cl₂:EtOAc; 100:0 to 70:30) afforded Ha-1 as an off-white powder(117.2 mg, 78%). ¹H NMR (CD₃OD, 200 MHz): δ 0.91 (t, J=7.1 Hz, 3H, H₂₁),1.62-1.35 (m, 4H, H₂₀ and H₁₉), 2.20 (s, 3H, H₁), 2.37 (t, J=6.8 Hz, 2H,H₁₈), 6.97 (ddd, J=8.0, 2.2, 0.9 Hz, 1H, H₉), 7.27-7.16 (m, 2H, H₄ andH₁₀), 7.39 (d, J=8.5 Hz, 1H, H₁₄), 7.57 (dt, J=7.8, 1.2 Hz, 1H, H₁₁),7.76-7.64 (m, 2H, H₇ and H₁₃). ¹³C NMR (CD₃OD, 50 MHz): δ 14.1, 19.8,22.7, 23.1, 31.9, 80.4, 95.4, 109.3, 120.2, 121.8, 123.6, 128.4 (2C),130.4, 130.5, 132.9 (2C), 137.2, 139.3, 139.7, 150.5, 159.6, 171.0.HRMS-ESI (m/z): [M+H]⁺ Calcd for C₂₃H₂₄N₃O₃S₂, 454.1254; Found:454.1249.

Product name Structure CRO32 (IIa-2)

N-(4-(3-((4-(oct-1-yn-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIa-2). The general procedure for the Sonogashira coupling was followedusing aryl halide 1a-3 (400.0 mg, 0.88 mmol), Pd(Ph₃)₄ (152.5 mg, 15%mol.), copper (I) iodide (25.1 mg, 15% mol.) and oct-1-yne (653.2 μL,4.43 mmol). Purification by silica gel flash chromatography(CH₂Cl₂:EtOAc; 100:0 to 70:30) afforded Ha-2 as an off-white powder(330.1 mg, 78%). ¹H NMR (CD₃OD, 200 MHz): δ 0.97-0.78 (m, 3H, H₂₃),1.62-1.15 (m, 9H, H₂₂, H₂₁, H₂₀ and H₁₉), 2.19 (s, 3H, H₁), 2.34 (t,J=6.8 Hz, 2H, H₁₈), 6.97 (ddd, J=8.0, 2.1 and 0.9 Hz, 1H, H₉), 7.26-7.14(m, 2H, H₄ and H₁₀), 7.38 (d, J=8.5 Hz, 2H, H₁₄), 7.56 (d, J=7.9 Hz, 1H,H₁₁), 7.74-7.65 (m, 3H, H₇ and H₁₃). ¹³C NMR (CD₃OD, 50 MHz): δ 14.5,20.2, 22.7, 23.7, 29.7, 29.8, 32.6, 80.4, 95.4, 109.3, 120.1, 121.7,123.6, 128.4 (2C), 130.3, 130.5, 132.9 (2C), 137.1, 139.3, 139.7, 150.4,159.6, 171.0. HRMS-ESI (m/z): [M+H]⁺ Calcd for C₂₅H₂₈N₃O₃S₂, 482.1567;Found: 482.1565.

Product name Structure AM305 (IIa-3)

N-(4-(3-((4-(3-hydroxyprop-1-yn-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIa-3). The general procedure for Sonogashira coupling was followedusing Ia-4 (300.0 mg, 0.66 mmol), Pd(PPh₃)₄ (114.4 mg, 15% mol.), copper(I) iodide (18.9 mg, 15% mol.) and propargyl alcohol (190.4 μL, 3.30mmol). Purification by silica gel flash chromatography (CH₂Cl₂:CH₃OH;100:0 to 95:5) afforded IIa-3 as a white-yellowish powder (201.9 mg,72%). ¹H NMR (CD₃OD, 200 MHz): δ 2.20 (s, 1H, H₁), 4.36 (s, 2H, H₁₈),6.97 (ddd, J=8.0, 2.2, 1.0 Hz, 1H, H₉), 7.34-7.14 (m, 2H, H₄ and H₁₀),7.51-7.42 (m, 2H, H₁₄), 7.63-7.53 (m, 1H, H₁₁), 7.78-7.66 (m, 3H, H₇ andH₁₃). ¹³C NMR (CD₃OD, 50 MHz): δ 22.7, 51.2, 84.0, 92.7, 109.3, 120.2,121.8, 123.7, 128.5 (2C), 129.1, 130.6, 133.1 (2C), 137.2, 139.3, 140.6,150.5, 159.6, 171.1. HRMS-ESI (m/z): [M+H]⁺ Calcd for C₂₀H₁₈N₃O₄S₂,428.0733; Found: 428.0736.

Product ref Structure AM311 (IIa-4)

N-(4-(3-((4-ethynylphenyl)sulfonamido)phenyl) thiazol-2-yl)acetamide(IIa-4). The general procedure for Sonogashira coupling was followedusing Ia-4 (300.0 mg, 0.66 mmol), PdP(Ph₃)₄ (114.4 mg, 15% mol.), copper(I) iodide (18.9 mg, 15% mol.) and trimethylsilylacetylene (281.9 μL,1.98 mmol). After completion of the reaction (about 2 h), the mixturewas filtered through a pad of silica (eluted with CH₂Cl₂/EtOAc 70:30)and the filtrate was concentrated under reduced pressure. The blackresidue was dissolved in CH₂Cl₂/CH₃OH (1:1; 2 mL:2 mL) and K₂CO₃ (456.1mg, 3.3 mmol) was added to the mixture. The resulting suspension wasstirred at r.t. for 5 h., adsorbed onto silica prior to purification bysilica gel flash chromatography (CH₂Cl₂/CH₃OH; 100:0 to 95:0) to affordthe pure titled compound as a yellow-brownish powder (88.2 mg, 34%). ¹HNMR (CD₃OD, 200 MHz): δ 2.20 (s, 3H, H₁), 3.70 (s, 1H, H₁₇), 6.97 (ddd,J=7.9, 2.3, 1.1 Hz, 1H, H₉), 7.30-7.14 (m, 2H, H₁₀ and H₄), 7.62-7.44(m, 3H, H₁₁ and H₁₄), 7.80-7.67 (m, 3H, H₇ and H₁₃). ¹³C NMR (CD₃OD, 50MHz): δ 22.7, 82.4, 82.9, 109.4, 120.2, 121.8, 123.7, 128.4 (3C), 130.6,133.6 (2C), 137.2, 139.2, 141.0, 150.4, 159.6, 171.0. HRMS-ESI (m/z):[M+H]⁺ Calcd for C₁₉H₁₆N₃O₃S₂, 398.0628; Found: 298.0626.

Product ref Structure AM366 (IIa-5)

N-(4-(3-((4-(cyclohexylethynyl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIa-5). The general procedure for Sonogashira coupling was followedusing Ia-4 (110 mg, 0.24 mmol), Pd(Ph₃)₂C₁₂ (25.7 mg, 15% mol.), copper(I) iodide (7.0 mg, 15% mol.) and cyclohexylacetylene (159.4 μL, 1.22mmol). Purification by silica gel flash chromatography(cyclohexane:EtOAc; 100:0 to 70:30) afforded IIa-5 as a white-yellowishpowder (55 mg, 47%). ¹H NMR (Acetone-d₆, 200 MHz): δ 1.58-1.22 (m, 6H,H₂₀ and H₂₁), 1.91-1.61 (m, 4H, H₁₉), 2.28 (s, 3H, H₁), 2.70-2.49 (m,1H, H₁₈), 7.09 (ddd, J=8.0, 2.3, 1.1 Hz, 1H, H₉), 7.26 (t, J=7.9 Hz, 1H,H₁₀), 7.38 (s, 1H, H₄), 7.48 (d, J=8.4 Hz, 2H, H₁₄), 7.61 (d, J=7.7 Hz,1H, H₁₁), 7.75 (d, J=8.5 Hz, 2H, H₁₃), 7.85 (t, J=1.9 Hz, 1H, H₇), 9.10(s, 1H, NH_(sulfonamide)), 11.17 (s, 1H, NH_(acetyl)). ¹³C NMR(Acetone-d₆, 50 MHz): δ 22.9, 25.4 (2C), 26.5, 30.3, 33.2 (2C), 80.3,99.0, 108.8, 119.6, 121.2, 123.1, 128.2 (2C), 129.7, 130.3, 132.7 (2C),136.8, 139.0, 139.6, 149.7, 159.0, 169.2. HRMS-ESI (m/z): [M+H]⁺ Calcdfor C₂₅H₂₆N₃O₃S₂, 480.1410; Found: 480.1413.

Product ref Structure CRO54 (IIa-6)

N-(3-(2-aminothiazol-4-yl)phenyl)-4-(oct-1-yn-1-yl)benzenesulfonamide(IIa-6). A suspension of IIa-2 in 2M aq. HCl/EtOH (1 mL/1 mL) wasstirred 2 h at 80° C., then 15 h at 50° C. The mixture was partitionedbetween EtOAc and saturated aq. Na₂CO₃, the aqueous phase was extractedwith EtOAc once. The combined organic layers were dried with Na₂SO₄ andconcentrated under reduced pressure to afford the pure titled compoundas a pale yellow solid (20.8 mg, 99%). R_(f)(EtOAc/cyclohexane, 1/1,v/v)=0.65. ¹H NMR (CD₃OD, 200 MHz): δ 1.00-0.77 (m, 3H, H₂₁), 1.73-1.18(m, 8H, H₁₇, H₁₈, H₁₉ and H₂₀), 2.39 (t, J=6.8 Hz, 2H, H₁₆), 6.73 (s,1H, H₂), 7.08-6.92 (m, 1H, H₈), 7.20 (t, J=8.1 Hz, 1H, H₈), 7.51-7.35(m, 4H, H_(aro)), 7.76-7.62 (m, 2H, H_(aro)). ¹³C NMR (CD₃OD, 50 MHz): δ14.4, 20.0, 23.6, 29.6, 29.7, 32.5, 90.3, 95.2, 103.5, 120.0, 121.4,123.5, 128.2 (2C), 130.2, 130.3, 132.8 (2C), 137.2, 139.1, 139.6, 150.9,171.3. HRMS-ESI (m/z): [M+H]⁺ Calcd for C₂₃H₂₆N₃O₂S₂, 440.1461; Found:440.1472.

Product ref Structure AM362 (IIa-7)

N-(3-(2-(methylamino)thiazol-4-yl)phenyl)-4-(oct-1-yn-1-yl)benzenesulfonamide(IIa-7). The general procedure for Sonogashira coupling was followedusing Ia-9 (325.0 mg, 0.77 mmol), Pd(PPh₃)₂Cl₂ (80.6 mg, 15% mol.),copper (I) iodide (21.9 mg, 15% mol.) and oct-1-yne (565.0 μL, 3.83mmol). Purification by silica gel flash chromatography(cyclohexane:EtOAc; 100:0 to 70:30) afforded IIa-7 as an off-whitepowder (80 mg, 23%). ¹H NMR (Acetone-d₆, 200 MHz): δ 0.85 (t, J=6.4 Hz,3H, H₂₂), 1.70-1.18 (m, 8H, H₁₈, H₁₉, H₂₀ and H₂₁), 2.40 (t, J=6.8 Hz,2H, H₁₇), 3.12-2.88 (m, 3H, H₁), 6.73 (s, 1H, NH_(thiazole)), 6.87 (s,1H, H₃), 7.11 (ddd, J=8.0, 2.2, 1.2 Hz, 1H, H₈), 7.23 (t, J=7.8 Hz, 1H,H₉), 7.53-7.44 (m, 2H, H₁₃), 7.58 (dt, J=7.7, 1.4 Hz, 1H, H₁₀),7.91-7.70 (m, 3H, H₆ and H₁₂), 9.10 (s, 1H, NH_(sulfonamide)). ¹³C NMR(Acetone-d₆, 50 MHz): δ 14.4, 19.8, 23.2, 29.2, 29.3, 31.8, 32.1, 80.2,95.2, 102.0, 119.5, 120.6, 123.0, 128.2 (2C), 129.6, 130.0, 132.7 (2C),137.4, 138.8, 139.7, 151.4, 170.7. HRMS-ESI (m/z): [M+H]⁺ Calcd forC₂₄H₂₈N₃O₂S₂, 454.1618; Found: 454.1615.

Product Structure AM369 (IIa-8)

4-(oct-1-yn-1-yl)-N-(3-(2-(phenylamino)thiazol-4-yl)phenyl)benzenesulfonamide(IIa-8). The general procedure for Sonogashira coupling was followedusing Ia-10 (100.0 mg, 0.21 mmol), Pd(PPh₃)₂Cl₂ (21.6 mg, 15% mol.),copper (I) iodide (5.9 mg, 15% mol.) and oct-1-yne (151.7 μL, 1.03mmol). Purification by silica gel flash chromatography(cyclohexane:EtOAc; 100:0 to 70:30) afforded IIa-8 as a brown powder (82mg, 75%). ¹H NMR (Acetone-d₆, 200 MHz): δ 0.86 (t, J=6.2 Hz, 3H, H₂₅),1.67-1.11 (m, 8H, H₂₁, H₂₂, H₂₃ and H₂₄), 2.39 (t, J=6.7 Hz, 2H, H₂₀),7.01 (t, J=7.3 Hz, 1H, H₁₂), 7.13 (d, J=9.1 Hz, 2H, H₆ and H₁₁), 7.33(dt, J=15.6, 7.8 Hz, 3H, H₁ and H₃), 7.48 (d, J=8.2 Hz, 2H, H₁₆), 7.66(d, J=7.7 Hz, 1H, H₁₃), 7.88-7.73 (m, 4H, H₂ and H₁₅), 7.92 (t, J=1.9Hz, 1H, H₉), 9.14 (s, 1H, NH_(thiazole)), 9.35 (s, 1H,NH_(sulfonamide)). ¹³C NMR (Acetone-d₆, 50 MHz): δ 14.4, 19.8, 23.2,29.2, 29.3, 32.1, 80.2, 95.3, 103.8, 118.3 (2C), 119.5, 120.8, 122.6,123.0, 128.2 (2C), 129.7, 129.9 (2C), 130.3, 132.7 (2C), 137.0, 139.0,139.7, 142.4, 151.3, 164.5. HRMS-ESI (m/z): [M+H]⁺ Calcd forC₂₉H₃₀N₃O₂S₂, 516.1774; Found: 516.1777.

Product ref Structure CRO47 (IIa-9)

N-(4-(3-((3-(oct-1-yn-1-Aphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIa-9). The general procedure for Sonogashira coupling was followedusing Ia-8 (113.0 mg, 0.25 mmol), Pd(PPh₃)₄ (44.0 mg, 15% mol.), copper(I) iodide (5.0 mg, 10% mol.) and oct-1-yne (185.0 μL, 1.25 mmol).Purification by silica gel flash chromatography (cyclohexane:EtOAc;100:0 to 60:40) afforded IIa-9 as a white-yellowish powder (103.0 mg,86%). R_(f) (EtOAc/cyclohexane, 1/1, v/v)=0.57. ¹H NMR (CD₃Cl, 200 MHz):δ 0.86 (t, J=6.0 Hz, 3H, H₂₅), 1.41-1.20 (m, 6H, H₂₂, H₂₃ and H₂₄),1.63-1.45 (m, 2H, H₂₁), 2.04 (s, 1H, H₁), 2.33 (t, J=6.9 Hz, 2H, H₂₀),7.00-6.85 (m, 2H, H_(aro)), 7.12 (t, J=8.0 Hz, 1H, H₁₀), 7.34-7.21 (m,2H, H_(aro)), 7.50-7.39 (m, 3H, H_(aro)), 7.61 (d, J=7.9 Hz, 2H,H_(aro)), 7.83 (s, 1H, H_(aro)), 10.51 (s, 1H, NH_(sulfonamide)). ¹³CNMR (CD₃Cl, 50 MHz): δ 14.3, 19.6, 22.7, 23.2, 28.7, 28.8, 31.5, 79.1,93.6, 108.9, 120.3, 121.7, 123.5, 125.8, 126.0, 129.2, 129.9, 130.3,135.7, 136.1, 136.9, 139.5, 148.7, 159.0, 168.6. HRMS-ESI (m/z): [M+H]⁺Calcd for C₂₅H₂₈N₃O₃S₂, 482.1567; Found: 482.1574.

Product ref Structure CRO49 (IIa-10)

N-(4-(3-((3-(3-hydroxyprop-1-yn-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIa-10). The general procedure for Sonogashira coupling was followedusing Ia-8 (113.0 mg, 0.25 mmol), Pd(PPh₃)₄ (44.0 mg, 15% mol.), copper(I) iodide (5.0 mg, 10% mol.) and propargyl alcohol (72.0 μL, 1.25mmol). Purification by silica gel flash chromatography(cyclohexane:EtOAc; 100:0 to 60:40) afforded Ha-10 as a white powder(76.1 mg, 71%), along with unreacted starting material X (22.0 mg, 20%).R_(f) (EtOAc/cyclohexane, 2/1, v/v)=0.38. ¹H NMR (CD₃OD, 200 MHz): δ2.21 (s, 3H, H₁), 4.37 (s, 2H, H₂₀), 6.99 (ddd, J=8.1, 2.3, 1.1 Hz, 1H,H₉), 7.29-7.17 (m, 2H, H₄ and H₁₀), 7.41 (t, J=7.8 Hz, 1H, H₁₆),7.62-7.51 (m, 2H, H_(aro)), 7.71 (tt, J=3.4, 1.5 Hz, 2H, H_(aro)), 7.80(t, J=1.8 Hz, 1H, H_(aro)). ¹³C NMR (CD₃OD, 50 MHz): δ 22.7, 51.2, 83.7,91.2, 109.3, 120.1, 121.7, 123.7, 125.5, 128.0, 130.4, 130.6, 131.0,136.7, 137.2, 139.2, 141.6, 150.4, 159.6, 171.1. HRMS-ESI (m/z): [M+H]⁺Calcd for C₂₀H₁₈N₃O₄S₂, 428.0733; Found: 428.0737.

Product ref Structure CRO50 (IIa-11)

N-(4-(3-((3-((trimethylsilyl)ethynyl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIa-11). The general procedure for Sonogashira coupling was followedusing Ia-8 (113.0 mg, 0.25 mmol), Pd(PPh₃)₄ (44.0 mg, 15% mol.), copper(I) iodide (5.0 mg, 10% mol.) and trimethylsilylacetylene (178.0 μL,1.25 mmol). Purification by silica gel flash chromatography(cyclohexane:EtOAc; 100:0 to 50:50) afforded IIa-11 as a pale yellowpowder (98.2 mg, 84%). R_(f) (EtOAc/cyclohexane, 1/1, v/v)=0.52. ¹H NMR(CD₃Cl, 200 MHz): δ 0.17 (s, 9H, H₂₀), 1.94 (s, 3H, H₁), 6.89 (d, J=7.1Hz, 2H, H_(aro)), 7.08 (t, J=7.8 Hz, 1H, H_(aro)), 7.46-7.17 (m, 3H,H_(aro)), 7.53 (d, J=7.5 Hz, 1H, H_(aro)), 7.66 (d, J=8.0 Hz, 1H,H_(aro)), 7.90 (s, 1H, H_(aro)), 8.18 (s, 1H, NH_(sulfonamide)), 10.97(s, 1H, NH_(acetyl)). ¹³C NMR (CD₃Cl, 50 MHz): δ 0.1 (3C), 23.0, 97.5,102.9, 109.0, 120.0, 121.5, 123.4, 124.7, 126.9, 129.3, 129.9, 130.6,135.6, 136.4, 136.9, 139.6, 148.6, 159.4, 169.0. HRMS-ESI (m/z): [M+H]⁺Calcd for C₂₂H₂₄N₃O₃S₂Si, 470.1023; Found: 470.1033.

Product ref Structure CRO53 (IIa-12)

N-(4-(3-((3-ethynylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIa-12). To a yellow solution of Ia-8 (51 mg, 0.108 mmol) in methanol(4 mL) was added solid K₂CO₃ (138 mg, 1.0 mmol). The resultingsuspension was stirred at r.t. for 14 h. The mixture was partitionedbetween EtOAc and water, the aqueous phase was extracted with EtOAconce. The combined organic layers were washed with brine, dried withNa₂SO₄ and concentrated under reduced pressure. Purification by silicagel flash chromatography (cyclohexane: EtOAc; 100:0 to 50:50) affordedIIa-12 as a white powder (41.8 mg, 97%). R_(f) (EtOAc/cyclohexane, 1/1,v/v)=0.47. ¹H NMR (CD₃OD, 200 MHz): δ 2.21 (s, 3H, H₁), 3.64 (s, 1H,H_(alkyne)), 6.99 (ddd, J=8.1, 2.3, 1.1 Hz, 1H, H₉), 7.33-7.16 (m, 2H,H₄ and H₁₀), 7.42 (t, J=7.8 Hz, 1H, H₁₆), 7.59 (ddt, J=7.8, 3.5, 1.4 Hz,2H, H_(aro)), 7.73 (dt, J=6.5, 1.6 Hz, 2H, H_(aro)), 7.85 (t, J=1.7 Hz,1H, H_(aro)). ¹³C NMR (CD₃OD, 50 MHz): δ 22.7, 81.1, 82.7, 109.3, 120.2,121.7, 123.7, 124.9, 128.4, 130.4, 130.6, 131.5, 137.2, 137.2, 139.2,141.6, 150.4, 159.6, 171.1. HRMS-ESI (m/z): [M+H]⁺ Calcd forC₁₉H₁₆N₃O₃S₂, 398.0628; Found: 398.0626.

Product ref Structure AM302 (IIIa-1)

N-(4-(3-((4-hexylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIIa-1). The reduction of IIa-1 was completed using a continuous flowapparatus. IIIa-1 (100.0 mg, 0.22 mmol) was dissolved in CH₃OH (HPLCgrade, 8 mL) and passed through a Pd/C (10%) cartridge at a flow of 0.5mL·min⁻¹, at 40° C. under 1 bar of H₂. The solvent was removed underreduced pressure to afford the pure titled compound as a white powder(87.3 mg, 87%). ¹H NMR (CD₃OD, 200 MHz): δ 0.86-0.80 (m, 3H, H₂₁),1.30-1.13 (m, 6H, H₁₈, H₁₉ and H₂₀), 1.60-1.43 (m, 2H, H₁₇), 2.20 (s,3H, H₁), 2.57 (t, J=7.6 Hz, 2H, H₁₆), 6.99 (ddd, J=7.9, 2.3, 1.1 Hz, 1H,H₉), 7.30-7.12 (m, 4H, H₄, H₁₀ and H₁₄), 7.69-7.53 (m, 4H, H₇, H₁₁ andH₁₃). ¹³C NMR (CD₃OD, 50 MHz): δ 14.4, 22.6, 23.6, 29.9, 32.1, 32.7,36.6, 109.0, 119.9, 121.6, 123.3, 128.3 (2C), 130.0 (2C), 130.3, 136.9,138.1, 139.4, 149.8, 150.4, 159.4, 170.9. HRMS-ESI (m/z): [M+H]⁺ Calcdfor C₂₃H₂₈N₃O₃S₂, 458.1572; Found: 458.1570.

Product ref Structure AM309 (IIIa-2)

(Z)—N-(4-(3-((4-(oct-1-en-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIIa-2). To a solution of IIa-2 (114.0 mg, 0.24 mmol) in CH₃OH(technical grade, 20 mL) was added Lindlar palladium (29.0 mg, 20% wt).The reaction mixture was allowed to react 2 h at r.t. and was filteredthrough a pad of Celite®. The resulting filtrate was concentrated underreduced pressure to afforded pure IIIa-2 as a white powder (86.7 mg,74%). ¹H NMR (CD₃OD, 200 MHz): δ 0.83 (t, J=6.2 Hz, 3H, H₂₃), 1.44-1.12(m, 8H, H₁₉, H₂₀, H₂₁ and H₂₂), 2.20 (s, 5H, H₁ and H₁₈) 5.74 (dt,J=11.8, 7.4 Hz, 1H, H₁₇), 6.37 (d, J=11.6 Hz, 1H, H₁₈), 7.09-6.90 (m,1H, H₉), 7.42-7.13 (m, 4H, H₁₀, H₄ and H₁₄), 7.57 (d, J=7.8 Hz, 1H,H₁₁), 7.78-7.62 (m, 3H, H₇ and H₁₃). ¹³C NMR (CD₃OD, 50 MHz): δ 14.5,22.7, 23.8, 29.7, 30.1, 30.8, 32.9, 109.2, 120.1, 121.8, 123.5, 128.3(2C), 128.6, 130.2 (2C), 130.5, 137.0, 137.1, 138.7, 139.5, 143.9,150.5, 159.6, 171.0. HRMS-ESI (m/z): [M+H]⁺ Calcd for C₂₅H₃₀N₃O₃S₂,484.1723; Found: 484.1729.

Product ref Structure AM295 (IIIa-3)

N-(4-(3-((4-octylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide(IIIa-3). The reduction of IIa-2 was completed using a continuous flowapparatus. IIa-2 (60.0 mg, 0.16 mmol) was dissolved in CH₃OH (HPLCgrade, 5 mL) and passed through a Pd/C (10%) cartridge at a flow of 0.5mL·min⁻¹, at 40° C. under 1 bar of H₂. The solvent was removed underreduced pressure to afford the pure titled compound as a white powder(35.2 mg, 60%). ¹H NMR (CD₃OD, 200 MHz): δ 0.85 (t, J=6.5 Hz, 3H, H₂₃),1.23 (d, J=11.3 Hz, 10H, H₂₂, H₂₁, H₂₀, H₂₉ and H₁₈), 1.53-1.43 (m, 2H,H₁₇), 2.20 (s, 3H, H₁), 2.55 (t, J=8.0 Hz, 2H, H₁₆), 6.99 (dd, J=7.5,1.6 Hz, 1H, H₉), 7.30-7.10 (m, 4H, H₄, H₁₀ and H₁₄), 7.60-7.49 (m, 1H,H₁₁), 7.74-7.60 (m, 3H, H₇ and H₁₃). ¹³C NMR (CD₃OD, 50 MHz): δ 14.6,22.7, 23.8, 30.3, 30.5, 30.6, 32.3, 33.1, 36.8, 109.2, 120.0, 121.7,123.4, 128.4 (2C), 130.1 (2C), 130.4, 137.0, 138.3, 139.6, 149.9, 150.5,159.5, 171.0. HRMS-ESI (m/z): [M+H]⁺ Calcd for C₂₅H₃₂N₃O₃S₂, 486.1884;Found: 486.1880.

Example 8: Anti-Cancer Activities of Compounds of the Invention Material& Methods Experimental Protocol for Assessment of Potency and EfficacyCell Cultures

Normal human melanocytes (NHM) prepared from foreskins of newborns weregrown under 5% CO₂ at 37° C. in MCDB 153 (Sigma) supplemented with 2%FCS, bovine pituitary extract (10 μg/ml), PMA (8 nM), bFGF (1 ng/ml),insulin (5 μg/ml), hydrocortisone (0.5 μg/ml), forskolin (10 μM),gentamicin (20 μg/ml), penicillin/streptomycin/amphotericin B (100 U/ml)(Invitrogen).

Normal human fibroblasts prepared from foreskins of newborns were grownunder 5% CO₂ at 37° C. in DMEM medium supplemented with 10% FCS andpenicillin/streptomycin (100 U/ml/50 mg/ml).

Different melanoma cell lines were purchased from American TissueCulture Collection (Molsheim, France). Cells were grown in RPMI 1640(A375, WM9 and patient melanoma cells) or in DMEM medium (Mel501)supplemented with 10% FCS and penicillin/streptomycin (100 U/ml/50mg/ml) at 37° C. and 5% CO₂.

Patient melanoma cells were prepared from biopsy after digestion for 1-2h with collagenase A (0.33 U/ml), dispase (0.85 U/ml) and Dnase I (144U/ml) at 37° C. Large debris were removed by filtration through a 70-mmcell strainer.

Trypan Blue Assays

Cells were seeded in 6-well plates (60000 cells/well), depleted andincubated with compounds for the times indicated. Then cells weredetached in the presence of 200 μl of HyQTase (Thermo) and 2 ml of RPMI1640 Glutamax (Gibco) was added to the cell solution. 10 μl of thissolution was stained for 1 minute with 10 μl of 0.4% trypan blue beforecounting with a Malassez chamber.

Western Blot assays

Proteins were extracted in Fisher buffer containing TRIS-HCl pH 7.5 50mM, NaCl 15 mM, Triton X-100 1% and proteases and phosphatasesinhibitors. Briefly, cell lysates (30 μg) were separated by SDS-PAGE,transferred onto a PVDF membrane (Millipore, Molsheim, France) and thenexposed to the appropriate antibodies. Proteins were visualized with theECL system from Amersham (Arlington, Heights, Ill., USA). The westernblots shown are representative of at least 3 independent experiments.

Results

FIG. 1: the Effect of Compounds Ia-7, IIIa-3, IIIa-1, IIIa-2, 11a-2,11a-9, Ia-4, Ha-11, IIa-12, IIa-6, Compared to Ia-1, on Cell Viabilityon A375 Melanoma Cells.Cell viability was assessed by measuring the number of cells alive insamples of different cells. The measure of cell viability as performedby cell counting using the trypan blue exclusion method. Results wereexpressed as the percentage of cells alive relatively to the number ofliving cells in the presence of DMSO, which corresponds to the negativecontrol associated to the 100% value.As depicted in FIG. 1, all new compounds displayed a better activity onA375 melanoma cells than the reference compound Ia-1 having a nudephenyl group. This clearly showed the benefit of introducing apolarsubstituents on this ring. A clear correlation between the size of theapolar group and the activity could be drawn and demonstrated having agreater impact on the viability than the substitution position (paraversus meta). The very active compounds IIa-2, IIIa-1 and IIIa-3substituted in para position respectively with an octyne, hexyl andoctyl chain, were selected to investigate their potential to promotecell death on resistant melanoma cell lines as well as other cancer celllines.FIG. 2a . Effect of Compounds 11a-2, IIIa-1 and IIIa-3 on Cell Viabilityon Melanoma Cells. PLX4032 was Used as a Positive Control.FIG. 2A shows a strong activity of compounds IIa-2, IIIa-1 and IIIa-3 at10 μM on both sensitive and resistant melanoma cell lines, contrary toreference B-Raf inhibitor drug PLX4032 which looses most activity onresistant A375 R5C3 an A375 RIV cell lines.FIG. 2b . Effect of Compounds IIa-2, IIIa-1 and IIIa-3 on Cell Viabilityon CML Cells. Imatinib was Used as a Positive Control.The activity of compounds IIa-2, IIIa-1 and IIIa-3 against CML cells wasalso evaluated. They showed a viability decrease in the same range thanthe standard BCR-ABL inhibitor Imatinib on sensitive K562 CML cells, andproved much more active on resistant K562 cells, displaying almost totalmortality for IIa-2 at this concentration.FIG. 2c . Effect of Compounds IIa-2, IIIa-1 and IIIa-3 on Cell Viabilityon Pancreatic Cancer Cells. Gemcitabine was Used as a Positive Control.The compounds IIa-2, IIIa-1 and IIIa-3 were next evaluated againstpancreatic cells, BxPC3 (sensitive) and MiaPACA cell lines (resistant tothe nucleoside analog Gemcitabine). All 3 compounds demonstrated a veryhigh activity on this cells, in the same range or better thanGemcitabine on sensitive cells. It is worth noting that they were stillvery active on resistant cells while Gemcitabine proved inactive. Thebest compound was again IIa-2 which induced total cell death for bothcell lines.

FIG. 3: Effect of Compound IIa-2 on the Viability of KHN and FHN CellLines.

Primary cell cultures of human normal melanocytes were prepared fromhuman foreskin. In order to determine the effect of compound IIa-2 oncell viability of melanocytes and fibroblasts, different concentrationof IIa-2 were added to the cell samples. The measure of cell viabilitywas performed in the same way as for FIG. 1. Results are expressed asthe percentage of living cells relatively to the number of living cellsin the presence of DMSO, as for FIG. 1.This figure showed that compound IIa-2 has no effect on normal cells(melanocytes and fibroblasts), which attested for its low toxicity andgood pharmacological profile

FIG. 4: Western Blots Showing the Mechanism of Death of Compound IIa-2(Apoptosis Versus Autophagy).

The Western blot in FIG. 4 shows the effect of IIa-2 (CRO32) on PARP(poly(ADP-ribose) polymerase) cleavage (apoptosis) and LCII/LCIIIconversion (autophagy). This figure clearly showed that compound IIa-2(10 μM) induced both apoptosis and autophagy cellular deaths in A375melanoma cell line. In contrast, only apoptosis is induced by PLX4032,which clearly attest for two different modes of action between IIa-2 andPLX4032. HA15, analogue of IIa-2 featuring a dansyl group, was used as apositive control and showed similar behavior as IIa-2 in this cell line.

REFERENCES

Throughout this application, various references describe the state ofthe art to which this invention pertains. The disclosures of thesereferences are hereby incorporated by reference into the presentdisclosure.

1-12. (canceled)
 13. A method of treating pancreas, prostate, breast andcolon cancers in a patient in need thereof, comprising administering tothe patient a therapeutically effective amount of a compound of generalformula (I)

wherein R₁ is selected from H, aryl and alkyl; R₂ is selected from H,alkyl, aryl and CO—R₆; R₃ is selected from H, halogen, alkyl, alkenyl,alkynyl, aryl, NHR₇, NR₇R₈, OR₇ and SR₇; R₄ is selected from (C₆-C₁₂)alkyl, (C₂-C₁₂) alkenyl, (C₂-C₁₂) alkynyl and (C₆-C₁₀) aryl, whereinaryl is unsubstituted or substituted by 1 to 5 alkyl groups, wherein thealkyl, alkenyl and alkynyl groups are linear, branched or cyclic andwherein the alkynyl group is optionally substituted with one to three OHgroups; R₅ is selected from H, R₆, aryl, OH, OR₆, O-aryl, SH, SR₆,S-aryl, CN, NO₂, CF₃, COOR₆, SO₂NR₇R₈, CONR₇R₈, NH₂, NHR₆, NH-aryl,NR₇R₈, NHCOR₆ and aminoacyl; R₆ is alkyl optionally substituted withhalogen, OH, SH, NH₂, O-alkyl, S-alkyl, NH-alkyl or NH-di(alkyl); R₇ andR₈ identical or different are H or alkyl optionally substituted withhalogen, OH, SH, NH₂, O-alkyl, S-alkyl, NH-alkyl or NH-di (alkyl); orpharmaceutically acceptable salts, isomers, tautomers, solvates orisotopic variations thereof;
 14. The method according to claim 13,wherein the cancer is pancreas cancer.
 15. The method according to claim13, wherein the cancer is prostate cancer.
 16. The method according toclaim 13, wherein the cancer is breast cancer.
 17. The method accordingto claim 13, wherein the cancer is colon cancer.
 18. The methodaccording to claim 13, wherein R₁ is H.
 19. The method according toclaim 13, wherein R₂ is selected from H, methyl or COCH₃.
 20. The methodaccording to claim 13, wherein R₃ is H.
 21. The method according toclaim 13, wherein R₄ is (C₆-C₁₂) alkyl, or (C₆-C₁₀) aryl, or CH═CHR₉,wherein R₉ is (C₁-C₁₂) alkyl, or C≡CR₁₀, wherein R₁₀ is selected from H,C₁-C₈ alkyl, hydroxy (C₁-C₈) alkyl, cyclo (C₃-C₈) alkyl andhydroxyl-cyclo (C₃-C₈ alkyl).
 22. The method according to claim 19,wherein (C₆-C₁₂) alkyl is hexyl, heptyl or octyl.
 23. The methodaccording to claim 19, wherein (C₆-C₁₀) aryl is phenyl, cyclopentyl, orcyclohexyl.
 24. The method according to claim 19, wherein R₉ is anhexyl.
 25. The method according to claim 13, wherein R₄ is in the metaor para position with respect to the sulfonyl group.
 26. The methodaccording to claim 13 wherein the compound is selected from:N-(4-(3-(4-(oct-1-ynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-(3-(oct-1-ynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-(3-(3-hydroxyprop-1-ynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-(3-((trimethylsilyl)ethynyl)phenylsulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-(3-ethynylphenylsulfonamido)phenyl)thiazol-2-yl)acetamide,N-(3-(2-aminothiazol-4-yl)phenyl)-4-(oct-1-ynyl)benzenesulfonamide,N-(3-(2-(methylamino)thiazol-4-yl)phenyl)-4-(oct-1-ynyl)benzenesulfonamide,4-(oct-1-ynyl)-N-(3-(2-(phenylamino)thiazol-4-yl)phenyl)benzenesulfonamide,N-(4-(3-((4-(cyclohexylethynyl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-([1,1′-biphenyl]-4-sulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-((2,3-dihydro-1H-indene)-5-sulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-((4′-methyl-[1,1′-biphenyl])-4-sulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-((4-octylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-((4-hexylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide,N-(4-(3-((4-(3-hydroxyprop-1-yn-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide,(Z)—N-(4-(3-((4-(oct-1-en-1-yl)phenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide,and N-(4-(3-((4-ethynylphenyl)sulfonamido)phenyl)thiazol-2-yl)acetamide.27. The method according to claim 13, wherein the patient is an animalor a human.